Macrocyclic tyrosine kinase inhibitor and uses thereof

ABSTRACT

The present invention belongs to the technical field of medicine, and relates to a macrocyclic tyrosine kinase inhibitor and the uses thereof. Specifically, the invention relates to a compound of general formula (I) or a pharmaceutically acceptable salt, ester or stereoisomer thereof, a preparation method therefor, a pharmaceutical formulation or a pharmaceutical composition containing same, and medical uses thereof.

TECHNICAL FIELD

The present invention belongs to the technical field of medicine, andparticularly relates to a macrocyclic tyrosine kinase inhibitor compoundor a pharmaceutically acceptable salt, ester or stereoisomer thereof.The tyrosine kinase is one or more of TRK, ALK and/or ROS1. The presentinvention further relates to a pharmaceutical composition or formulationcontaining the compound or the pharmaceutically acceptable salt, esteror stereoisomer thereof, a method for preparing the compound or thepharmaceutically acceptable salt, ester or stereoisomer thereof, anduses of the compound or the pharmaceutically acceptable salt, ester orstereoisomer thereof.

BACKGROUND

Cancer, also known as malignant tumor, is a serious threat to humanhealth and life. In 2004, 7.4 million people died from cancer worldwide.In 2008, the third national survey on causes of death in China showsthat the cancer death rate in China has increased by about 80% in thelast 30 years, and nearly 2 million people died from cancer every year,which lead to a very severe situation.

Molecular targeted therapy has been a major breakthrough in cancertreatment in recent years. Compared with traditional treatment meanssuch as surgery, radiotherapy and chemotherapy, the molecular targetedtherapy breaks new ground for cancer treatment with its high specificityand relatively low toxic side effects, and gradually serves as astandard treatment scheme for patients with advanced cancer. Proteinkinases, a large field of targeted therapy, are key regulators for cellgrowth, proliferation and survival, and both genetic and epigeneticalterations may lead to the occurrence of cancer.

Anaplastic lymphoma kinase (ALK) was named after its first discovery inthe anaplastic large cell lymphoma AMS3 cell line. EML4 belongs toechinoderm microtubule-associated protein family, and consists of anN-terminus base region, a hydrophobic echinoderm microtubule-associatedprotein region and a WD repeat region. It has been reported that anEML4-ALK fusion gene is related to the formation of tumors, and anN-terminus base region plays the most important role. Soda et al. (2007)reported the EML4-ALK gene fusion in non-small cell lung cancer (NSCLC)for the first time which is a result of inversion of the short arm ofchromosome 2, with EML4 fused to an intracellular ALK kinase region atthe N-terminus. EML4-ALK fusion sites are various, and at least 8EML4-ALK mutants are formed. ALK mutations have been found in a varietyof cancers, including anaplastic large cell lymphoma (ALCL), non-smallcell lung cancer, inflammatory myofibroblast tumor, colorectal cancer,breast cancer and several others.

ROS1 is also a tyrosine kinase receptor that has attracted muchattention. ROS1 is located at region 6q21 and the full length cDNAthereof contains 44 exons. ROS1 encodes 2347 amino acids with amolecular weight of 259 kDa. The basic structure of ROS1 consists of anextramembrane region (amino acids 1-1861), a transmembrane region (aminoacids 1862-1882) and a tyrosine kinase active region (amino acids1883-2347) within the membrane. The first proto-oncogene fusion site ofROS1 (FIG-ROS1) is found in glioblastoma, and an intermediate deletionof 240 bases on 6q21 results in the expression of FIG-ROS1 fusionprotein, which activates the activity of tyrosine kinase. ROS1 mutationshave also been found in a variety of cancers, including glioblastoma,non-small cell lung cancer, colorectal cancer, breast cancer and thelike.

Trks are a family of nerve growth factor-activated tyrosine kinases,including three subtypes TrkA, TrkB and TrkC, and are encoded by NTRK1(neuronal receptor tyrosine kinase 1), NTRK2 and NTRK3 genes,respectively. After Trk kinases are phosphorylated, downstream signalmolecules can be activated, thereby playing roles in regulating cellproliferation, differentiation, metabolism, apoptosis and the like. TheNTRK gene can be fused with other genes, which causes the highexpression of Trk kinases or continuous increase of the activity of Trkkinases, and finally may lead to cancer. NTRK gene fusion occurs in avariety of adult and pediatric solid tumors, including breast cancer,colorectal cancer, non-small cell lung cancer and various sarcomas.

The above three tyrosine kinases have strong homology. The ROS1 gene andthe ALK gene have 49% homology in the tyrosine kinase region sequence,and have a high 77% homology in ATP binding site of kinase catalyticregion. The kinase region sequences of TRK A/B/C have over 80% homology.The TRK A gene, the ROS1 gene and the ALK gene have about 40% homologyin the tyrosine kinase region sequence. The marketed ALK inhibitorCrizotinib has ROS1 and TRK inhibitory activities simultaneously, andthe TRK inhibitor Entrectinib also has ALK and ROS1 inhibitoryactivities.

At present, both the marketed ALK/ROS1 inhibitors and the NTRKinhibitors filed NDA in 2017 have appeared drug resistance duringlong-term medication, mainly because the amino acid sequence of thekinase protein is changed due to gene mutation, such as common mutationsites in ALK kinases including L1196M, L1152R, G1202R, G1269A, 1151Tins,S1206Y, C1156Y, F1174L and the like, common mutation sites in ROS1kinases including G2032R, D2033N, S1986F, L2026M, L1951R and the like,and common mutation sites in NTRK kinases including G595R, G623R, G667C,G623E, L686M and the like. Therefore, the development of an anti-tumordrug which has strong efficacy and low toxicity and can solve drugresistance has very important clinical value and significance.

SUMMARY OF THE INVENTION

In order to solve the aforementioned problem, the present inventionprovides a compound with a novel structure and a tyrosine kinasereceptor inhibition effect or a pharmaceutically acceptable salt, esteror stereoisomer thereof. The tyrosine kinase receptor may be one or moreof TRK, ALK and ROS1. In addition, the compound has better inhibitoryactivity on one or more of TrkA, TrkB and TrkC kinases, and has higherexposure and bioavailability in organisms.

In order to solve the aforementioned problem, the present invention alsoprovides a method for treating a cancer disease mediated by one or moretyrosine kinase receptors of TRK, ALK and ROS1 using the compound of thepresent invention, or the pharmaceutically acceptable salt, ester orstereoisomer thereof. The cancer disease may have developed resistanceto one or more existing therapeutically active agents, and theresistance may be caused by a mutation in the gene encoding targets.Preferably, the mutant gene is an NTRK gene.

In one aspect, the present application provides a compound of generalformula (I) or a pharmaceutically acceptable salt, ester or stereoisomerthereof,

wherein:

M¹, M², M³, M⁴, M⁵, M⁶ and M⁷ are each independently selected from C,C(R¹) and N;

X¹, X², X³ and L¹, when present, are each independently selected from—C(R⁵)(R⁶)—, —N(R⁴)—, —O—, —S—, —S(O)— and —S(O)₂—;

ring A is selected from 3-10 membered cycloalkyl, 3-10 memberedheterocyclyl, 6-8 membered monoaryl, 8-10 membered fused aryl, 5-10membered monoheteroaryl and 8-10 membered fused heteroaryl;

ring B is absent or selected from 3-10 membered cycloalkyl and 3-10membered heterocyclyl, and when ring B is absent, X² and X³ are directlyconnected by a chemical bond;

R¹, R², R³, R⁵ and R⁶, when present, are each independently selectedfrom hydrogen, halogen, nitro, cyano and the following groups optionallysubstituted with one or more Q¹: C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,—OR^(a), —SR^(a), —N^(a)R^(b), —C(O)R^(a), —C(O)OR^(a), —OC(O)R^(a),—OC(O)OR^(a), —OC(O)NR^(a)R^(b), —C(O)NR^(a)R^(b), —NR^(a)C(O)R^(b),—NR^(a)C(O)OR^(b), —NR^(a)C(O)NR^(a)R^(b), —S(O)R^(a), —S(O)OR^(a),—OS(O)R^(a), —OS(O)OR^(a), —OS(O)NR^(a)R^(b), —S(O)NR^(a)R^(b),—NR^(a)S(O)R^(b), —NR^(a)S(O)OR^(b), —NR^(a)S(O)NR^(a)R^(b),—S(O)₂R^(a), —S(O)₂OR^(a), —OS(O)₂R^(a), —OS(O)₂OR^(a),—OS(O)₂NR^(a)R^(b), —S(O)₂NR^(a)R^(b), —NR^(a)S(O)₂R^(b),—NR^(a)S(O)₂OR^(b), —NR^(a)S(O)₂NR^(a)R^(b), 3-10 membered cycloalkyl,3-10 membered heterocyclyl, 6-10 membered aryl and 5-10 memberedheteroaryl; Q¹, when present, is independently selected from hydroxy,amino, halogen, nitro, cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ alkylamino,di(C₁₋₆ alkyl)amino, halo C₁₋₆ alkyl, hydroxy C₁₋₆ alkyl, amino C₁₋₆alkyl, halo C₁₋₆ alkoxy, 3-10 membered cycloalkyl, 3-10 memberedheterocyclyl, 6-10 membered aryl and 5-10 membered heteroaryl;

R⁴, when present, is independently selected from hydrogen and thefollowing groups optionally substituted with one or more Q²: C₁₋₆ alkyl,C₂₋₆ alkenyl, C₂₋₆ alkynyl, —C(O)R^(a), —C(O)OR^(a), —C(O)NR^(a)R^(b),—S(O)R^(a), —S(O)OR^(a), —S(O)NR^(a)R^(b), —S(O)₂R^(a), —S(O)₂OR^(a),—S(O)₂NR^(a)R^(b), 3-10 membered cycloalkyl, 3-10 membered heterocyclyl,6-10 membered aryl and 5-10 membered heteroaryl; Q², when present, isindependently selected from hydroxy, amino, halogen, nitro, cyano, C₁₋₆alkyl, C₁₋₆ alkoxy, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino, halo C₁₋₆alkyl, hydroxy C₁₋₆ alkyl, amino C₁₋₆ alkyl, halo C₁₋₆ alkoxy, 3-10membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl and5-10 membered heteroaryl;

or X¹, together with L¹, forms 3-10 membered cycloalkyl, 3-10 memberedheterocyclyl or 5-6 membered monoheteroaryl; and/or L¹, together withsome ring atoms of ring A, forms 3-10 membered cycloalkyl or 3-10membered heterocyclyl; and/or X², together with some ring atoms of ringA, forms 3-10 membered cycloalkyl or 3-10 membered heterocyclyl; the3-10 membered cycloalkyl, 3-10 membered heterocyclyl and 5-6 memberedmonoheteroaryl are optionally each independently substituted with asubstituent selected from: R⁴, R⁵, R⁶, halogen, amino, hydroxy, nitro,cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino,halo C₁₋₆ alkyl, hydroxy C₁₋₆ alkyl, amino C₁₋₆ alkyl and halo C₁₋₆alkoxy;

R^(a) and R^(b), when present, are each independently selected fromhydrogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino,halo C₁₋₆ alkyl, hydroxy C₁₋₆ alkyl, amino C₁₋₆ alkyl, halo C₁₋₆ alkoxy,3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryland 5-10 membered heteroaryl; and

m, n, p, q and t are each independently selected from 0, 1, 2, 3, 4 and5.

In some embodiments, the present application provides a compound of theaforementioned general formula (I) or a pharmaceutically acceptablesalt, ester or stereoisomer thereof, wherein:

M¹, M², M³, M⁴, M⁵, M⁶ and M⁷ are each independently selected from C,C(R¹) and N, at least one of which is N;

X¹, X², X³ and L¹, when present, are each independently selected from—C(R⁵)(R⁶)—, —N(R⁴)— and —O—;

ring A is selected from 6-8 membered monoaryl and 5-8 memberedmonoheteroaryl;

ring B is absent or selected from 3-8 membered cycloalkyl and 3-8membered heterocyclyl;

R¹, R², R³, R⁵ and R⁶, when present, are each independently selectedfrom hydrogen, halogen, nitro, cyano and the following groups optionallysubstituted with 1 to 3 Q¹: C₁₋₆ alkyl, —OR^(a), —NR^(a)R^(b),—C(O)R^(a), —C(O)OR^(a), —OC(O)R^(a), —OC(O)OR^(a), —OC(O)NR^(a)R^(b),—C(O)NR^(a)R^(b), —NR^(a)C(O)R^(b), —NR^(a)C(O)OR^(b),—NR^(a)C(O)NR^(a)R^(b), 3-8 membered cycloalkyl, 3-8 memberedheterocyclyl, 6-8 membered aryl and 5-8 membered heteroaryl; Q¹, whenpresent, is independently selected from hydroxy, amino, halogen, nitro,cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino,halo C₁₋₆ alkyl, hydroxy C₁₋₆ alkyl, amino C₁₋₆ alkyl and halo C₁₋₆alkoxy;

R⁴, when present, is independently selected from hydrogen and thefollowing groups optionally substituted with 1 to 3 Q²: C₁₋₆ alkyl,—C(O)R^(a), —C(O)OR^(a), —C(O)NR^(a)R^(b), 3-8 membered cycloalkyl and3-8 membered heterocyclyl; Q², when present, is independently selectedfrom hydroxy, amino, halogen, nitro, cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy,C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino, halo C₁₋₆ alkyl, hydroxy C₁₋₆alkyl, amino C₁₋₆ alkyl, halo C₁₋₆ alkoxy, 3-8 membered cycloalkyl and3-8 membered heterocyclyl;

or X¹, together with L¹, forms 3-8 membered cycloalkyl, 3-8 memberedheterocyclyl or 5-6 membered monoheteroaryl; and/or L¹, together withsome ring atoms of ring A, forms 3-8 membered cycloalkyl or 3-8 memberedheterocyclyl; and/or X², together with some ring atoms of ring A, forms3-8 membered cycloalkyl or 3-8 membered heterocyclyl; the 3-8 memberedcycloalkyl, 3-8 membered heterocyclyl and 5-6 membered monoheteroarylare optionally each independently substituted with a substituentselected from: R⁴, R⁵, R⁶, halogen, amino, hydroxy, nitro, cyano, C₁₋₆alkyl, C₁₋₆ alkoxy, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino, halo C₁₋₆alkyl, hydroxy C₁₋₆ alkyl, amino C₁₋₆ alkyl and halo C₁₋₆ alkoxy;

R^(a) and R^(b), when present, are each independently selected fromhydrogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino,halo C₁₋₆ alkyl, hydroxy C₁₋₆ alkyl, amino C₁₋₆ alkyl, halo C₁₋₆ alkoxy,3-8 membered cycloalkyl and 3-8 membered heterocyclyl; and

m, n, p, q and t are each independently selected from 0, 1, 2, 3, 4 and5.

In some embodiments, the present application provides a compound of theaforementioned general formula (I) or a pharmaceutically acceptablesalt, ester or stereoisomer thereof, wherein:

M¹, M², M³, M⁴, M⁵, M⁶ and M⁷ are each independently selected from C,C(R¹) and N, at least one of which is N;

X¹ and L¹ are each independently selected from —C(R⁵)(R⁶)— and —N(R⁴)—;

X² and X³, when present, are each independently selected from—C(R⁵)(R⁶)—, —N(R⁴)— and —O—;

ring A is selected from phenyl and 5-6 membered monoheteroaryl;

ring B is absent or selected from 3-6 membered monocycloalkyl and 3-6membered monoheterocyclyl;

R¹, R², R³, R⁵ and R⁶, when present, are each independently selectedfrom hydrogen, halogen, nitro, cyano and the following groups optionallysubstituted with 1 to 3 Q¹: C₁₋₆ alkyl, —OR^(a), —NR^(a)R^(b),—C(O)R^(a), —C(O)OR^(a), —C(O)NR^(a)R^(b), 3-6 membered cycloalkyl, 3-6membered heterocyclyl, phenyl and 5-6 membered heteroaryl; Q¹, whenpresent, is independently selected from hydroxy, amino, halogen, nitro,cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino,halo C₁₋₆ alkyl, hydroxy C₁₋₆ alkyl, amino C₁₋₆ alkyl and halo C₁₋₆alkoxy;

R⁴, when present, is independently selected from hydrogen and thefollowing groups optionally substituted with 1 to 3 Q²: C₁₋₆ alkyl,—C(O)R^(a), 3-6 membered cycloalkyl and 3-6 membered heterocyclyl; Q²,when present, is selected from hydroxy, amino, halogen, nitro, cyano,C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino, halo C₁₋₆alkyl, hydroxy C₁₋₆ alkyl, amino C₁₋₆ alkyl, halo C₁₋₆ alkoxy, 3-6membered cycloalkyl and 3-6 membered heterocyclyl;

or X¹, together with L¹, forms 3-6 membered monoheterocyclyl or 5-6membered monoheteroaryl; and/or L¹, together with some ring atoms ofring A, forms 3-6 membered monocycloalkyl or 3-6 memberedmonoheterocyclyl; and/or X², together with some ring atoms of ring A,forms 3-6 membered monocycloalkyl and 3-6 membered monoheterocyclyl; the3-6 membered monoheterocyclyl, 5-6 membered monoheteroaryl and 3-6membered monocycloalkyl are optionally each independently substitutedwith a substituent selected from: R⁴, R⁵, R⁶, halogen, amino, hydroxy,C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino, halo C₁₋₆alkyl, hydroxy C₁₋₆ alkyl, amino C₁₋₆ alkyl and halo C₁₋₆ alkoxy;

R^(a) and R^(b), when present, are each independently selected fromhydrogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino,halo C₁₋₆ alkyl, hydroxy C₁₋₆ alkyl, amino C₁₋₆ alkyl, halo C₁₋₆ alkoxy,3-6 membered cycloalkyl and 3-6 membered heterocyclyl;

m is 1, 2 or 3;

n, p and q are each independently selected from 0, 1, 2 and 3; and

t is 0 or 1.

In some embodiments, the present application provides a compound of theaforementioned general formula (I) or a pharmaceutically acceptablesalt, ester or stereoisomer thereof, wherein the compound is of thestructure shown as general formula (II),

wherein:

M², M⁵ and M⁶ are each independently selected from CH and N; and

X¹, X², X³, L¹, A, R², m, n, p and q are described as above for generalformula (I).

In some embodiments, the present application provides a compound of theaforementioned general formula (I) or a pharmaceutically acceptablesalt, ester or stereoisomer thereof, wherein the compound is of thestructure shown as general formula (III),

wherein:

ring C is selected from 3-6 membered saturated monoheterocyclyl and 5-6membered nitrogen-containing monoheteroaryl, preferably 5-6 memberedsaturated monoheterocyclyl; the 3-6 membered saturated monoheterocyclyland 5-6 membered nitrogen-containing monoheteroaryl are optionally eachindependently substituted with a substituent selected from: R⁴, R⁵, R⁶,halogen, amino, hydroxy, C₁₋₄ alkyl, C₁₋₄ alkoxy, C₁₋₄ alkylamino,di(C₁₋₄ alkyl)amino, halo C₁₋₄ alkyl, hydroxy C₁₋₄ alkyl, amino C₁₋₄alkyl and halo C₁₋₄ alkoxy;

—(X²)_(p)— is selected from —C(R⁵)(R⁶)—, —C(R⁵)(R⁶)—C(R⁵)(R⁶)—,—N(R⁴)—C(R⁵)(R⁶)—, —O—C(R⁵)(R⁶)—, —C(R⁵)(R⁶)—N(R⁴)— and —C(R⁵)(R⁶)—O—,and the left chemical bond thereof is connected to ring A and the rightchemical bond thereof is connected to X³;

—(X³)_(q)— is selected from —C(R⁵)(R⁶)—, —C(R⁵)(R⁶)—C(R⁵)(R⁶)—,—N(R⁴)—C(R⁵)(R⁶)—, —O—C(R⁵)(R⁶)—, —C(R⁵)(R⁶)—N(R⁴)— and —C(R⁵)(R⁶)—O—,and the left chemical bond thereof is connected to X² and the rightchemical bond thereof is connected to carbonyl;

ring A is selected from phenyl and 5-6 membered monoheteroaryl,preferably 5-6 membered nitrogen-containing heteroaryl;

R², when present, is independently selected from hydrogen, halogen andthe following groups optionally substituted with 1 to 3 Q¹: C₁₋₄ alkyl,—OR^(a) and —NR^(a)R^(b); Q¹, when present, is independently selectedfrom hydroxy, amino, halogen, nitro, cyano, C₁₋₄ alkyl, C₁₋₄ alkoxy,C₁₋₄ alkylamino, di(C₁₋₄ alkyl)amino, halo C₁₋₄ alkyl, hydroxy C₁₋₄alkyl, amino C₁₋₄ alkyl and halo C₁₋₄ alkoxy;

R^(a) and R^(b), when present, are each independently selected fromhydrogen and C₁₋₄ alkyl;

R⁵ and R⁶, when present, are each independently selected from hydrogen,halogen, hydroxy, amino, C₁₋₄ alkyl and C₁₋₄ alkoxy;

R⁴, when present, is independently selected from hydrogen and C₁₋₄ alkyloptionally substituted with 1 to 2 Q²; Q², when present, isindependently selected from hydroxy, amino, halogen and C₁₋₄ alkoxy;

n is 0, 1 or 2; and

M¹, M⁵ and M⁶ are described as above for general formula (I) or (II).

In some embodiments, the present application provides a compound of theaforementioned general formula (III) or a pharmaceutically acceptablesalt, ester or stereoisomer thereof, wherein ring C is optionallysubstituted 5-6 membered saturated monoheterocyclyl, more preferablyoptionally substituted 5-6 saturated nitrogen-containing heterocyclyl.

In some embodiments, the present application provides a compound of theaforementioned general formula (III) or a pharmaceutically acceptablesalt, ester or stereoisomer thereof, wherein the compound is of thestructure shown as general formula (IV),

wherein each optional substituent on ring C is independently selectedfrom R⁴, R⁵, R⁶, halogen, amino, hydroxy, nitro, cyano, C₁₋₆ alkyl, C₁₋₆alkoxy, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino, halo C₁₋₆ alkyl, hydroxyC₁₋₆ alkyl, amino C₁₋₆ alkyl and halo C₁₋₆ alkoxy.

In some embodiments, the present application provides a compound of theaforementioned general formula (III) or a pharmaceutically acceptablesalt, ester or stereoisomer thereof, wherein:

ring C is selected from aziridinyl, azetidinyl, pyrrolidinyl,imidazolidinyl, pyrazolidinyl, piperidinyl, morpholinyl, piperazinyl,hexahydropyridazinyl, hexahydropyrimidinyl, pyrrolyl, imidazolyl,pyrazolyl, pyridinyl or pyrimidinyl; each of which is optionallysubstituted with a substituents selected from: fluorine, chlorine,bromine, iodine, hydroxy, amino, methyl, ethyl, propyl, isopropyl,butyl, isobutyl, sec-butyl, methoxy, ethoxy, propoxy, isopropoxy,methylamino, dimethylamino, ethylamino, diethylamino, trifluoromethyland trifluoromethoxy;

ring A is selected from phenyl, pyrrolyl, imidazolyl, pyrazolyl,thiazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl,1,2,4-triazolyl, 1,2,3-triazolyl, 1,3,4-oxadiazolyl, 1,2,4-oxadiazolyl,1,3,4-thiadiazolyl, 1,2,4-thiadiazolyl, pyridinyl, pyrimidinyl,pyridazinyl, 1,3,5-triazinyl and tetrazinyl;

R², when present, is independently selected from hydrogen, fluorine,chlorine, bromine, iodine, hydroxy, amino, methyl, ethyl, propyl,isopropyl, butyl, isobutyl, sec-butyl, methylamino, dimethylamino,ethylamino, diethylamino, methoxy, ethoxy, propoxy, isopropoxy,trifluoromethyl and trifluoromethoxy;

R⁵ and R⁶, when present, are each independently selected from hydrogen,fluorine, chlorine, bromine, iodine, hydroxy, amino, methyl, ethyl,propyl, isopropyl, butyl, isobutyl, sec-butyl, methoxy, ethoxy, propoxyand isopropoxy; and

R⁴, when present, is independently selected from hydrogen, methyl,ethyl, propyl, isopropyl, butyl, isobutyl and sec-butyl.

In some embodiments, the present application provides a compound of theaforementioned general formulas (I)-(IV) or a pharmaceuticallyacceptable salt, ester or stereoisomer thereof, wherein:

ring A is selected from

and preferably the wavy line marked with “1” represents the connectionpoint of ring A to L¹ and the wavy line marked with “2” represents theconnection point of ring A to X²;

—(X²)_(p)— is selected from —C(R⁵)(R⁶)—, —C(R⁵)(R⁶)—C(R⁵)(R⁶)—,—N(R⁴)—C(R⁵)(R⁶)— and —O—C(R⁵)(R⁶)—, and preferably the left chemicalbond thereof is connected to ring A and the right chemical bond thereofis connected to X³; and

—(X³)_(q)— is selected from —C(R⁵)(R⁶)—, —C(R⁵)(R⁶)—N(R⁴)— and—C(R⁵)(R⁶)—O—, and preferably the left chemical bond thereof isconnected to X² and the right chemical bond thereof is connected tocarbonyl.

In some embodiments, the present application provides a compound of theaforementioned general formula (I) or a pharmaceutically acceptablesalt, ester or stereoisomer thereof, wherein the compound is of thestructure shown as general formula (V),

In some embodiments, the present application provides a compound of theaforementioned general formula (I) or a pharmaceutically acceptablesalt, ester or stereoisomer thereof, wherein:

X¹ is —N(R⁴)—;

L¹ is —C(R⁵)(R⁶)—;

ring A is selected from phenyl and 5-6 membered nitrogen-containingheteroaryl;

R², when present, is independently selected from hydrogen, halogen andthe following groups optionally substituted with 1 to 3 Q¹: C₁₋₄ alkyl,—OR^(a) and —NR^(a)R^(b); Q¹, when present, is independently selectedfrom hydroxy, amino, halogen, nitro, cyano, C₁₋₄ alkyl, C₁₋₄ alkoxy,C₁₋₄ alkylamino, di(C₁₋₄ alkyl)amino, halo C₁₋₄ alkyl, hydroxy C₁₋₄alkyl, amino C₁₋₄ alkyl and halo C₁₋₄ alkoxy;

R^(a) and R^(b), when present, are each independently selected fromhydrogen and C₁₋₄ alkyl;

R⁵ and R⁶, when present, are each independently selected from hydrogen,halogen, hydroxy, amino, C₁₋₄ alkyl and C₁₋₄ alkoxy;

R⁴, when present, is independently selected from hydrogen and C₁₋₄ alkyloptionally substituted with 1 to 2 Q²; Q², when present, isindependently selected from hydroxy, amino, halogen and C₁₋₄ alkoxy; and

n is 0, 1 or 2.

In some embodiments, the present application provides a compound of theaforementioned general formula (I) or a pharmaceutically acceptablesalt, ester or stereoisomer thereof, wherein:

ring A is selected from phenyl, pyrrolyl, imidazolyl, pyrazolyl, furyl,thienyl, thiazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl,1,2,4-triazolyl, 1,2,3-triazolyl, 1,3,4-oxadiazolyl, 1,2,4-oxadiazolyl,1,3,4-thiadiazolyl, 1,2,4-thiadiazolyl, pyridinyl, pyrimidinyl,pyridazinyl, 1,3,5-triazinyl and tetrazinyl;

R², when present, is independently selected from hydrogen, fluorine,chlorine, bromine, iodine, hydroxy, amino, methyl, ethyl, propyl,isopropyl, butyl, isobutyl, sec-butyl, methylamino, dimethylamino,ethylamino, diethylamino, methoxy, ethoxy, propoxy, isopropoxy,trifluoromethyl and trifluoromethoxy;

R⁵ and R⁶, when present, are each independently selected from hydrogen,fluorine, chlorine, bromine, iodine, hydroxy, amino, methyl, ethyl,propyl, isopropyl, butyl, isobutyl, sec-butyl, methoxy, ethoxy, propoxyand isopropoxy; and

R⁴, when present, is independently selected from hydrogen, methyl,ethyl, propyl, isopropyl, butyl, isobutyl and sec-butyl.

In some embodiments, the present application provides a compound of theaforementioned general formula (I) or a pharmaceutically acceptablesalt, ester or stereoisomer thereof, wherein:

ring A is selected from

and preferably the wavy line marked with “1” represents the connectionpoint of ring A to L¹ and the wavy line marked with “2” represents theconnection point of ring A to X²;

—(X²)_(p)— is selected from —C(R⁵)(R⁶)—, —C(R⁵)(R⁶)—C(R⁵)(R⁶)—,—N(R⁴)—C(R⁵)(R⁶)— and —O—C(R⁵)(R⁶)—, and preferably the left chemicalbond thereof is connected to ring A and the right chemical bond thereofis connected to X³; and

—(X³)_(q)— is selected from —C(R⁵)(R⁶)—, —C(R⁵)(R⁶)—N(R⁴)— and—C(R⁵)(R⁶)—O—, and preferably the left chemical bond thereof isconnected to X² and the right chemical bond thereof is connected tocarbonyl.

Any substituent and any optional group thereof in the aforementionedtechnical solution described in the present application can be combinedwith each other to form a new and complete technical solution, and theformed new technical solution is encompassed within the scope of thepresent invention.

In some embodiments, the present application provides a compound of theaforementioned general formula (I) or a pharmaceutically acceptablesalt, ester or stereoisomer thereof, wherein the compound is selectedfrom:

No. Compound Compound 1

Compound 2

Compound 2-1

Compound 3

Compound 3-1

Compound 4

Compound 5

Compound 6

Compound 7

Compound 8

Compound 9

Compound 10

Compound 11

Compound 11-1

Compound 12

Compound 13

Compound 13-1

Compound 14

Compound 15

Compound 16

Compound 17

* represents that the carbon atom is a chiral carbon atom having asingle configuration (R configuration or S configuration).

In some embodiments, the present application provides a compound of theaforementioned general formula (I) or a pharmaceutically acceptablesalt, ester or stereoisomer thereof, wherein the compound is selectedfrom:

No. Compound Compound 2′

Compound 3′

Compound 11′

In another aspect, the present application provides a pharmaceuticalformulation comprising a compound of the aforementioned general formula(I), (II), (III), (IV) or (V), or a pharmaceutically acceptable salt,ester or stereoisomer thereof, and one or more pharmaceuticallyacceptable carriers and/or excipients. The pharmaceutical formulationmay be any pharmaceutically acceptable dosage form. Pharmaceuticallyacceptable excipients are substances that are non-toxic, compatible withactive ingredient, and otherwise biologically compatible with organism.The choice of a particular excipient will depend on an administrationmode used to treat a particular patient or disease type and state.Examples of pharmaceutically acceptable excipient include, but are notlimited to, solvents, diluents, dispersant, suspending agents,surfactants, isotonizing agent, thickening agents, emulsifiers, binders,lubricants, stabilizers, hydrating agents, emulsification accelerators,buffers, absorbents, colorants, ion exchangers, mold release agents,coating agents, flavoring agents, antioxidants and the like which areconventional in the pharmaceutical field. If necessary, perfumingagents, preservatives, sweeteners and the like may be further added tothe pharmaceutical composition.

In some embodiments, the above pharmaceutical formulations may beadministered to a patient or subject in need of such treatment via oral,parenteral, rectal, or pulmonary administration and the like. For oraladministration, the pharmaceutical composition may be prepared into oralformulations, for example conventional oral solid formulations such astablets, capsules, pills, granules and the like; or can also be preparedinto oral liquid formulations such as oral solutions, oral suspensions,syrups and the like. When the pharmaceutical compositions are preparedinto oral formulations, suitable fillers, binders, disintegrants,lubricants and the like may be added. For parenteral administration, theabove pharmaceutical formulations may also be prepared into injectionformulations, including injections, sterile powders for injection andconcentrated solutions for injection. The injection formulations can beprepared by a conventional method known in the pharmaceutical field, andduring the preparation, no additive may be added, or suitable additivesmay be added according to the properties of the drug. For rectaladministration, the pharmaceutical compositions may be prepared intosuppositories and the like. For pulmonary administration, thepharmaceutical compositions may be prepared into inhalationformulations, aerosols, powder spray or spray and the like.

In another aspect, the present application also provides apharmaceutical composition comprising a compound of the aforementionedgeneral formula (I), (II), (III), (IV) or (V), or a pharmaceuticallyacceptable salt, ester or stereoisomer thereof, and one or more secondtherapeutically active agents for use in combination with the tyrosinekinase inhibitor compounds of the present application in the treatmentand/or prevention of tyrosine kinase-mediated diseases and relatedconditions, such as pain, cancer, inflammation, neurodegenerativediseases, autoimmune diseases, infectious diseases and the like.

The pain may be of any origin or etiology, including but not limited toone or more of inflammatory pain, visceral pain, cancer-induced pain,chemotherapy pain, wound pain, surgical and postoperative pain, laborpain, acute pain, chronic pain, intractable pain, somatic pain,nociceptive pain, neuropathic pain, blood-borne pain, immunogenetricpain, endocrine-derived pain, metabolic-induced pain, cardiogenic pain,headache, phantom limb pain and dental pain. Therapeutically activeagents suitable for use in combination therapy for pain include, but arenot limited to, Nav1.7 channel modulators, opioid analgesics,non-steroidal anti-inflammatory drugs, sedatives,selective/non-selective cyclooxygenase inhibitors, antiepileptics,antidepressants, local anesthetics, 5-HT receptor blockers, 5-HTreceptor agonists, ergot alkaloids, β-receptor blockers, M receptorblockers, nitrates, vitamin K and the like.

The cancer includes, but is not limited to, one or more of lung cancer,colon cancer, prostate cancer, breast cancer, liver cancer, lymphoma,thyroid cancer, multiple myeloma, soft tissue sarcoma, ovarian cancer,cervical cancer, fallopian tube carcinoma, renal cell carcinoma, gastriccancer, gastrointestinal stromal tumor, bone cancer, basal cellcarcinoma, peritoneal cancer, dermatofibroma, pancreatic cancer,esophageal cancer, glioblastoma, head and neck cancer, inflammatorymyofibroblast tumor and anaplastic large cell lymphoma. Secondtherapeutically active agents suitable for use in combination therapyfor cancer include, but are not limited to, mitotic inhibitors,alkylating agents, antimetabolites, antisense DNA or RNA, antitumorantibiotics, growth factor inhibitors, signal transduction inhibitors,cell cycle inhibitors, enzyme inhibitors, retinoid receptor modulators,proteasome inhibitors, topoisomerase inhibitors, biological responsemodifiers, hormones, angiogenesis inhibitors, cell growth inhibitors,targeted antibodies, HMG-CoA reductase inhibitors, prenyl-proteintransferase inhibitors and the like.

The inflammation includes, but is not limited to, atherosclerosis,allergy, and inflammation due to infection or injury. Therapeuticallyactive agents suitable for use in combination therapy for inflammationinclude, but are not limited to, steroidal anti-inflammatory drugs andnon-steroidal anti-inflammatory drugs.

The neurodegenerative disease includes, but is not limited to, one ormore of Alzheimer's disease, Parkinson's disease, amyotrophic lateralsclerosis and Huntington's disease. Therapeutically active agentssuitable for use in the combination therapy for neurodegenerativediseases include, but are not limited to, dopamine-mimetics, dopaminereceptor agonists, agents affecting dopamine metabolism, NMDA receptorantagonists, adenosine A_(2A) receptor inhibitors, agents affecting DArelease and reuptake, central anticholinergics, cholinesteraseinhibitors, 5-HT agonists, α2 adrenergic receptor antagonists,antidepressants, cholinergic receptor agonists, β/γ secretaseinhibitors, H3 receptor antagonists or antioxidant agents and the like.

The autoimmune disease includes, but is not limited to, one or more ofrheumatoid arthritis, Sjogren's syndrome, type I diabetes and lupuserythematosus. Therapeutically active agents suitable for use in thecombination therapy for autoimmune diseases include, but are not limitedto, antirheumatic drugs, non-steroidal anti-inflammatory drugs,glucocorticoid drugs, TNF antagonists, cyclophosphamide, mycophenolatemofetil, cyclosporine and the like, which is used for alleviating thedisease.

The infectious diseases include trypanosomiasis and the like.

In some embodiments, the pharmaceutical compositions further compriseone or more pharmaceutically acceptable excipients described as above.

In some embodiments, the compound of general formula (I), (II), (III),(IV) or (V), or the pharmaceutically acceptable salt, ester orstereoisomer thereof, and the second therapeutically active agent may bepresent in one formulation, i.e., as a combined formulation, or may bepresent in separate formulations simultaneously or sequentiallyadministrating to a subject.

In another aspect, the present application also relates to uses of thecompound of the aforementioned general formula (I), (II), (III), (IV) or(V), or the pharmaceutically acceptable salt, ester or stereoisomerthereof in the preparation of medications for the treatment and/orprevention of diseases and related conditions mediated by one or moretyrosine kinases of TRK, ALK and/or ROS1.

In some embodiments, the diseases and related conditions mediated by oneor more tyrosine kinases of TRK, ALK and/or ROS1 include, but are notlimited to, pain, cancer, inflammation, neurodegenerative disease,autoimmune disease, infectious disease and the like. The cancerincludes, but is not limited to, lung cancer, colon cancer, rectalcancer, prostate cancer, breast cancer, liver cancer, gallbladdercancer, cholangiocarcinoma, leukemia, melanoma, lymphoma, thyroidcancer, multiple myeloma, soft tissue sarcoma, ovarian cancer, cervicalcancer, fallopian tube carcinoma, renal cell carcinoma, gastric cancer,gastrointestinal stromal tumor, bone cancer, basal cell carcinoma,peritoneal cancer, dermatofibroma, pancreatic cancer, esophageal cancer,glioblastoma, head and neck cancer, inflammatory myofibroblast tumor,anaplastic large cell lymphoma or neuroblastoma and the like. The pain,inflammation, neurodegenerative disease, autoimmune disease andinfectious disease are defined as above.

In some embodiments, the lung cancer includes small cell lung cancer andnon-small cell lung cancer. In some embodiments, the non-small cell lungcancer comprises lung adenocarcinoma, squamous cell carcinoma and largecell carcinoma.

In some embodiments, the cancers mediated by one or more tyrosinekinases of TRK, ALK and/or ROS1 include a cancer that is at leastpartially resistant to one or more existing target therapeuticallyactive agents.

In some embodiments, the cancer resistance is caused by one or moremutations in the gene encoding TRK, ALK and/or ROS1 kinase receptors.

In some embodiments, the mutation sites of the ALK target are located atL1196M, L1152R, G1202R, G1269A, G1269S, 1151Tins, S1206R, S1206Y/C,C1156Y, F1174L, F1174S, R1050H, F1245C/I/L/V, R1275L/Q, T1151M, M1166R,I1170N, I1170S, I1171N, I1171T, I1171S, V1180L, I1183T, L1196M, A1200V,L1204F, L1240V, D1270G, Y1278S, R1192P, G1128A, G1286R, T1343I, D1203N,E1210K, F1174S, F1174C/L/V, F1245C/L, L1252R, G1296M/Q, T1151K/M, V1180Land the like in polypeptide. In some embodiments, the mutation sites ofthe ROS1 target are located at G2032R, D2033N, S1986F, L2026M, L1951R,L2155S, G2101A, K2003I and the like in polypeptide. In some embodiments,the mutation sites of the TRK target are located at G517R, A542V, V573M,F589L, F589C, G595R, G595S, D596V, F600L, F646V, C656Y, C656F, L657V,G667S, G667C, Y676S, G623R, G667C, G623E, L686M, G545R, A570V, Q596E,Q596P, V601G, F617L, F617C, G623S, D624V, R630K, C682Y, C682F, L683V,G693S, G713S, C685F, C685Y, L686V, G696A, G639R and the like inpolypeptide.

In some embodiments, the compound of the present application may also beused to treat diseases and related conditions mediated by tyrosinekinases selected from JAK2, SRC, FYN, LYN, YES, FGR, FAK, ARK5 or anycombination thereof, preferably the diseases are cancers.

In another aspect, the present application also provides a method fortreating diseases and related conditions mediated by one or moretyrosine kinases of TRK, ALK and ROS1, comprising administering to apatient in need thereof an effective amount of compound of theaforementioned general formula (I), (II), (III), (IV) or (V) or thepharmaceutically acceptable salt, ester or stereoisomer thereof, theaforementioned pharmaceutical formulation, or the aforementionedpharmaceutical composition, wherein the diseases and related conditionsmediated by one or more tyrosine kinases of TRK, ALK and ROS1 aredescribed as above.

The “effective amount” refers to a dosage of a drug that is capable ofreducing, delaying, inhibiting or curing the condition in a subject. Theamount of the administered dose is determined by factors such as theadministration mode of drugs, the pharmacokinetics of agents, theseverity of diseases, and the individual signs (sex, weight, height,age) of the subject.

In another aspect, the present application also provides a method forpreparing a compound of general formula (I), comprising the steps asfollows:

intermediate 1 and intermediate 2 are subjected to a plurality ofreactions such as substitution reaction, deprotection reaction,acylation reaction (using an acylation reagent) and the like in thepresence of an organic solvent and a catalyst at a proper temperature toobtain the compound of the general formula (I).

The substituents and variables in the above formulas are defined asabove.

The organic solvent includes a common reaction solvent known to thoseskilled in the art, which may be a polar solvent or a non-polar solvent,a protic solvent or a non-protic solvent, and preferably a polarnon-protic solvent such as dichloromethane, trichloromethane,tetrahydrofuran, acetonitrile, dimethylformamide, dimethyl sulfoxide andthe like.

The catalyst includes a dehydration catalyst and a base catalyst.Examples of the dehydration catalyst include diethyl azodicarboxylate,azobisformyl dipiperidine, tributylphosphine and the like. The basecatalyst includes an organic base and an inorganic base, preferably anorganic base, more preferably an organic amine bases such astriethylamine, pyridine, N,N-diisopropylethylamine and the like.

Examples of the substitution reaction include the reaction between acidsand alcohols to make esters, dehydration between alcohols to makeethers, and dehydration between carboxylic acids to make acidanhydrides.

The deprotection reaction may be, for example, a reaction for removinghydroxy or amino protecting groups.

The acylation reaction may be, for example, a reaction between acylationagents and amino or hydroxy to make amides or esters.

The intermediates involved in the preparation method of the presentapplication are either commercially available or self-made, and can beprepared by those skilled in the art according to known conventionalchemical reactions, and the preparation method thereof is also withinthe scope of the present application.

In the present application, all compounds are named according tochemical structures thereof, and if a compound name is not consistentwith chemical structure thereof for the same compound, the chemicalstructure shall prevail.

In the present application, unless otherwise specified, scientific andtechnical terms used herein have the meanings generally understood bythose skilled in the art, however, in order to better understand thepresent invention, definitions of some terms are provided below. If thedefinitions of terms provided herein are not consistent with themeanings generally understood by those skilled in the art, thedefinitions and explanations of the terms provided in the presentapplication shall prevail.

The “halogen” described herein refers to fluorine, chlorine, bromine oriodine.

The “C₁₋₆ alkyl” described herein refers to a straight-chain or branchedalkyl having 1 to 6 carbon atoms, and includes, for example, “C₁₋₅alkyl”, “C₁₋₄ alkyl”, “C₁₋₃ alkyl”, “C₁₋₂ alkyl”, “C₂₋₆ alkyl”, “C₂₋₅alkyl”, “C₂₄ alkyl”, “C₂₋₃ alkyl”, “C₃₋₆ alkyl”, “C₃₋₅ alkyl”, “C₃₋₄alkyl”, “C₄₋₆ alkyl”, “C₄₋₅ alkyl”, “C₅₋₆ alkyl” and the like. Examplesof C₁₋₆ alkyl include, but are not limited to, methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl,isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl,3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl,2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl,1,3-dimethylbutyl, 2,3-ethylbutyl, 2-ethylbutyl, 1,2-dimethylpropyl andthe like. The “C₁₋₄ alkyl” described herein refers to a specific examplehaving 1 to 4 carbon atoms in the C₁₋₆ alkyl.

The “C₁₋₆ alkoxy” described herein refers to “C₁₋₆ alkyl-O-”, wherein“C₁₋₆ alkyl” is defined as above. The “C₁₋₄ alkoxy” described hereinrefers to “C₁₋₄ alkyl-O-”, wherein “C₁₋₄ alkyl” is defined as above.

The “C₂₋₆ alkenyl” described herein refers to a straight-chain, branchedor cyclic alkenyl having at least one double bond and 2 to 6 carbonatoms, and includes, for example, “C₂₋₅ alkenyl”, “C₂₄ alkenyl”, “C₂₋₃alkenyl”, “C₃₋₆ alkenyl”, “C₃₋₅ alkenyl”, “C₃₋₄ alkenyl”, “C₄₋₆alkenyl”, “C₄₋₅ alkenyl”, “C₅₋₆ alkenyl” and the like. Examples of C₂₋₆alkenyl include, but are not limited to, ethenyl, 1-propenyl,2-propenyl, 1-butenyl, 2-butenyl, 1,3-butadienyl, 1-pentenyl,2-pentenyl, 3-pentenyl, 1,3-pentadienyl, 1,4-pentadienyl, 1-hexenyl,2-hexenyl, 3-hexenyl, 1,4-hexadienyl, cyclopentenyl,1,3-cyclopentadienyl, cyclohexenyl, 1,4-cyclohexadienyl and the like.

The “hydroxy C₁₋₆ alkyl”, “amino C₁₋₆ alkyl” and “halo C₁₋₆ alkyl”described herein each refer to the group wherein the hydrogen of C₁₋₆alkyl group is substituted by one or more hydroxy, amino or halogengroups, and wherein the C₁₋₆ alkyl is defined as above.

The “halo C₁₋₆ alkoxy” described herein refers to C₁₋₆ alkoxy groupwherein the hydrogen is substituted by one or more halogen groups.

The “C₁₋₆ alkylamino” and “di(C₁₋₆ alkyl)amino” described herein referto C₁₋₆ alkyl-NH— and

respectively.

The “ring B is absent” described herein refers to X² directly connectedwith X³ by a single chemical bond.

The “3-10 membered cycloalkyl” includes “3-8 membered cycloalkyl” and“8-10 membered fused cycloalkyl”.

The “3-8 membered cycloalkyl” described herein refers to a saturated orpartially saturated and non-aromatic monocyclic cycloalkyl having 3 to 8ring carbon atoms, and includes “3-8 membered saturated cycloalkyl” and“3-8 membered partially saturated cycloalkyl”, and preferably “3-4membered cycloalkyl”, “3-5 membered cycloalkyl”, “3-6 memberedcycloalkyl”, “3-7 membered cycloalkyl”, “4-5 membered cycloalkyl”, “4-6membered cycloalkyl”, “4-7 membered cycloalkyl”, “4-8 memberedcycloalkyl”, “5-6 membered cycloalkyl”, “5-7 membered cycloalkyl”, “5-8membered cycloalkyl”, “6-7 membered cycloalkyl”, “6-8 memberedcycloalkyl”, “7-8 membered cycloalkyl”, “3-6 membered saturatedcycloalkyl”, “5-8 membered saturated cycloalkyl”, “5-7 memberedsaturated cycloalkyl”, “5-6 membered saturated cycloalkyl” and the like.Specific examples of “3-8 membered saturated cycloalkyl” include, butare not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, cyclooctyl and the like; specific examples of the “3-8membered partially saturated cycloalkyl” include, but are not limitedto, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl,cyclohexenyl, cyclohex-1,3-dienyl, cyclohex-1,4-dienyl, cycloheptenyl,cyclohept-1,3-dienyl, cyclohept-1,4-dienyl, cyclohept-1,3,5-trienyl,cyclooctenyl, cyclooct-1,3-dienyl, cyclooct-1,4-dienyl,cyclooct-1,5-dienyl, cyclooct-1,3,5-trienyl, cyclooctatetraenyl and thelike.

The “8-10 membered fused cycloalkyl” described herein refers to asaturated or partially saturated and non-aromatic cycloalkyl having 8 to10 ring carbon atoms, which is formed by two or more cyclic structuressharing two adjacent carbon atoms (one ring of the fused cycloalkyl maybe an aromatic ring, but the fused cycloalkyl as a whole isnon-aromatic), and includes “8-9 membered fused cycloalkyl”, “9-10membered fused cycloalkyl” and the like, which may be fused in a manneras 5-6 membered cycloalkano 5-6 membered cycloalkyl, benzo 5-6 memberedcycloalkyl and the like. Examples of 8-10 membered fused cycloalkylinclude, but are not limited to, bicyclo[3.1.0]hexyl,bicyclo[4.1.0]heptyl, bicyclo[2.2.0]hexyl, bicyclo[3.2.0]heptyl,bicyclo[4.2.0]octyl, octahydro cyclopentadienyl, octahydro-1H-indenyl,decahydronaphthyl, tetradecahydrophenanthryl, bicyclo[3.1.0]hex-2-enyl,bicyclo[4.1.0]hept-3-enyl, bicyclo[3.2.0]hept-3-enyl,bicyclo[4.2.0]oct-3-enyl, 1,2,3,3a-tetrahydrocyclopentadienyl,2,3,3a,4,7,7a-hexahydro-1H-indenyl, 1,2,3,4,4a,5,6,8a-octahydronaphthyl,1,2,4a,5,6,8a-hexahydronaphthyl,1,2,3,4,5,6,7,8,9,10-decahydrophenanthryl, benzocyclopentyl,benzocyclohexyl, benzocyclohexenyl, benzocyclopentenyl and the like.

The “3-10 membered heterocyclyl” described herein includes “3-8 memberedheterocyclyl” and “8-10 membered fused heterocyclyl”.

The “3-8 membered heterocyclyl” described herein refers to a saturatedor partially saturated and non-aramatic monocyclic cyclic group having 3to 8 ring atoms, which contains at least one (e.g., 1, 2, 3, 4 or 5)ring heteroatom(s), and the heteroatoms are nitrogen atoms, oxygen atomsand/or sulfur atoms. Optionally, a ring atom (e.g., a carbon atom, anitrogen atom, or a sulfur atom) may be substituted by an oxo group(forming, for example, a C═O, N═O, S═O or SO₂ ring member). The “3-8membered heterocyclyl” includes “3-8 membered saturated heterocyclyl”and “3-8 membered partially saturated heterocyclyl”. Preferably, the“3-8 membered heterocyclyl” contains 1 to 3 heteroatoms such as one ortwo heteroatoms selected from nitrogen and oxygen atoms, or one nitrogenatom. Preferably, the “3-8 membered heterocyclyl” is “3-7 memberedheterocyclyl”, “3-6 membered heterocyclyl”, “4-7 membered heterocyclyl”,“4-6 membered heterocyclyl”, “6-8 membered heterocyclyl”, “5-7 memberedheterocyclyl”, “5-6 membered heterocyclyl”, “3-6 membered saturatedheterocyclyl”, “5-6 membered saturated heterocyclyl”, “3-6 memberednitrogen-containing heterocyclyl”, “3-6 membered saturatednitrogen-containing heterocyclyl”, “5-6 membered nitrogen-containingheterocyclyl”, “5-6 membered saturated nitrogen-containing heterocyclyl”and the like. The “3-8 membered heterocyclyl” may, for example, containsonly one or two nitrogen atoms, or contains one nitrogen atom and one ortwo other heteroatoms (e.g., oxygen atoms and/or sulfur atoms). Specificexamples of “3-8 membered heterocyclyl” include, but are not limited to,aziridinyl, 2H-aziridinyl, diazaziridinyl, 3H-diazacyclopropenyl,azetidinyl, 1,4-dioxanyl, 1,3-dioxanyl, 1,3-dioxolanyl, 1,4-dioxadienyl,tetrahydrofuryl, dihydropyrrolyl, pyrrolidinyl, imidazolidinyl,4,5-dihydroimidazolyl, pyrazolidinyl, 4,5-dihydropyrazolyl,2,5-dihydrothienyl, tetrahydrothienyl, 4,5-dihydrothiazolyl,thiazolidinyl, piperidinyl, tetrahydropyridinyl, piperidinonyl,tetrahydropyridinonyl, dihydropiperidinonyl, piperazinyl, morpholinyl,4,5-dihydrooxazolyl, 4,5-dihydroisoxazolyl, 2,3-dihydroisoxazolyl,oxazolidinyl, 2H-1,2-oxazinyl, 4H-1,2-oxazinyl, 6H-1,2-oxazinyl,4H-1,3-oxazinyl, 6H-1,3-oxazinyl, 4H-1,4-oxazinyl, 4H-1,3-thiazinyl,6H-1,3-thiazinyl, 2H-pyranyl, 2H-pyran-2-onyl, 3,4-dihydro-2H-pyranyland the like.

The “8-10 membered fused heterocyclyl” described herein refers to asaturated or partially saturated and non-aromatic cyclic group having 8to 10 ring atoms and with at least one of the ring atom being aheteroatom (one ring of the fused heterocyclyl may be an aromatic ring,but the fused heterocyclyl as a whole is non-aromatic), which is formedby two or more cyclic structures sharing two adjacent atoms with eachother, and the heteroatoms are nitrogen atoms, oxygen atoms and/orsulfur atoms. Optionally, a ring atom (e.g., a carbon atom, a nitrogenatom, or a sulfur atom) may be substituted by an oxo group (forming, forexample, a C═O, N═O, S═O or SO₂ ring member). The “8-10 membered fusedheterocyclyl” includes “8-9 membered fused heterocyclyl”, “9-10 memberedfused heterocyclyl” and the like, which may be fused in a manner as 5-6membered heterocyclo 5-6 membered heterocyclyl, 5-6 membered heterocyclo5-6 membered cycloalkyl, benzo 5-6 membered heterocyclyl and 5-6membered heteroaro 5-6 membered heterocyclyl, wherein 5-6 memberedheteroaryl is defined as below. Examples of “8-10 membered fusedheterocyclyl” include, but are not limited to, pyrrolidinocyclopropyl,cyclopentazacyclopropyl, pyrrolidinocyclobutyl, pyrrolidinopyrrolidinyl,pyrrolidinopiperidinyl, pyrrolidinopiperazinyl, pyrrolidinomorpholinyl,piperidinomorpholinyl, benzopyrrolidinyl,tetrahydroimidazo[4,5-c]pyridinyl, 3,4-dihydroquinazolinyl,1,2-dihydroquinoxalinyl, benzo[d][1,3]dioxolyl,1,3-dihydroisobenzofuranyl, 2H-chromenyl, 2H-chromen-2-onyl,4H-chromenyl, 4H-chromen-4-onyl, chromanyl, 4H-1,3-benzoxazinyl,4,6-dihydro-1H-furo[3,4-d]imidazolyl,3a,4,6,6a-tetrahydro-1H-furo[3,4-d]imidazolyl,4,6-dihydro-1H-thieno[3,4-d]imidazolyl,4,6-dihydro-1H-pyrrolo[3,4-d]imidazolyl, alkyl benzimidazolyl,octahydrobenzo [d]imidazolyl, decahydroquinolinyl,hexahydrothienoimidazolyl, hexahydrofuroimidazolyl,4,5,6,7-tetrahydro-1H-benzo[d]imidazolyl,octahydrocyclopenta[c]pyrrolyl, dihydroindolyl, dihydroisoindolyl,benzoxazolylalkyl, benzothiazolylalkyl, 1,2,3,4-tetrahydroisoquinolinyl,1,2,3,4-tetrahydroquinolinyl, 4H-1,3-benzoxazinyl and the like.

The “6-10 membered aryl” described herein includes “6-8 memberedmonocycloaryl” and “8-10 membered fused ring aryl”.

The “6-8 membered monocycloaryl” described herein refers tomonocycloaryl having 6 to 8 ring carbon atoms, examples of whichinclude, but are not limited to, phenyl, cyclooctatetraenyl and thelike, and preferably phenyl.

The “8-10 membered fused ring aryl” described herein refers to anaromatic cyclic group having 8 to 10 ring carbon atoms and formed by twoor more cyclic structures sharing two adjacent carbon atoms with eachother, and preferably “9-10 membered fused ring aryl”, such as naphthyland the like.

The “5-10 membered heteroaryl” described herein includes “5-8 memberedsingle monoheteroaryl” and “8-10 membered fused heteroaryl”.

The “5-8 membered monoheteroaryl” described herein refers to an aromaticmonocyclic cyclic group having 5 to 8 ring atoms (at least one of whichis a heteroatom such as a nitrogen atom, an oxygen atom or a sulfuratom). Optionally, a ring atom (e.g., a carbon atom, a nitrogen atom, ora sulfur atom) may be substituted by an oxo group (forming, for example,a C═O, N═O, S═O or SO₂ ring member). “5-8 membered monoheteroaryl”includes “5-7 membered monoheteroaryl”, “5-6 membered monoheteroaryl”,“5-6 membered nitrogen-containing monoheteroaryl”, “6 memberednitrogen-containing monoheteroaryl” and the like, wherein at least oneof the ring heteroatoms of the “nitrogen-containing monoheteroaryl” is anitrogen atom, for example, it may contain only one or two nitrogenatoms, or one nitrogen atom and one or two other heteroatoms (e.g.,oxygen atoms and/or sulfur atoms), or two nitrogen atoms and one or twoother heteroatoms (e.g., oxygen atoms and/or sulfur atoms). Specificexamples of “5-8 membered monoheteroaryl” include, but are not limitedto, furyl, thienyl, pyrrolyl, thiazolyl, isothiazolyl, thiadiazolyl,oxazolyl, isoxazolyl, oxadiazolyl, imidazolyl, pyrazolyl,1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl,1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, pyridyl, 2-pyridonyl, 4-pyridonyl,pyrimidinyl, pyridazinyl, pyrazinyl, 1,2,3-triazinyl, 1,3,5-triazinyl,1,2,4,5-tetrazinyl, azacycloheptyltrienyl, 1,3-diazacycloheptyltrienyl,azacyclooctatetraenyl and the like. Specific examples of “5-6 memberedmonoheteroaryl” include, but are not limited to, those having 5 to 6ring atoms in the specific examples of the above-mentioned “5-8 memberedmonoheteroaryl”.

The “8-10 membered fused heteroaryl” described herein refers to anaromatic cyclic structure, which is formed by two or more cyclicstructures sharing two adjacent atoms with each other, and contains 8 to10 ring atoms (at least one of which is a heteroatom such as a nitrogenatom, an oxygen atom or a sulfur atom). Optionally, a ring atom (e.g., acarbon atom, a nitrogen atom, or a sulfur atom) may be substituted by anoxo group (forming, for example, a C═O, N═O, S═O or SO₂ ring member).“8-10 membered fused heteroaryl” includes “9-10 membered fusedheteroaryl”, “8-9 membered fused heteroaryl” and the like, which may befused in a manner as benzo 5-6 membered heteroaryl, 5-6 memberedheteroaro 5-6 membered heteroaryl and the like. Specific examples of“8-10 membered fused heteroaryl” include, but are not limited to,pyrrolopyrrolyl, pyrrolofuryl, pyrazolopyrrolyl, pyrazolothienyl,furothienyl, pyrazoloxazolyl, benzofuryl, benzoisofuryl, benzothienyl,indolyl, isoindolyl, benzoxazolyl, benzoimidazolyl, indazolyl,benzotriazolyl, quinolinyl, 2-quinolinonyl, 4-quinolinonyl,1-isoquinolinonyl, isoquinolinyl, acridinyl, phenanthridinyl,benzopyridazinyl, phthalazinyl, quinazolinyl, quinoxalinyl, purinyl,naphthyridinyl and the like.

The term “optionally substituted by . . . ” described herein includesboth “substituted by . . . ” and “not substituted by . . . ”.

The “pharmaceutically acceptable salt” described herein refers to a saltformed by an acidic functional group (e.g., —COOH, —OH, —SO₃H and thelike) in the compound combining with a suitable inorganic or organicbase (including alkali metal salts, alkaline-earth metal salts, ammoniumsalts and salts with nitrogen-containing organic bases) or a salt formedby a basic functional group (e.g., —NH₂ and the like) in the compoundcombining with a suitable inorganic or organic acid (e.g., carboxylicacid and the like).

The “stereoisomer” described herein means that when the compound of thepresent invention can exists in a form of a racemate or a racemicmixture, a single enantiomer, a mixture of diastereoisomer or a singlediastereoisomer as the compound contains one or more asymmetric centers.The compound of the present invention may have asymmetric centers andthus result in the presence of two optical isomers. The scope of thepresent invention includes all possible optical isomers and mixturesthereof. If the compound of the present invention contains an olefinicdouble bond, the scope of the present invention includes cis-isomer andtrans-isomer unless otherwise specified. The compounds described hereinmay exist in tautomeric (one of the functional group isomers) formshaving different connection points of hydrogen through one or moredouble bond shifts, for example, a ketone and enol form thereof areketo-enol tautomers. All tautomers and the mixture thereof are includedwithin the scope of the present invention. All enantiomers,diastereoisomers, racemates, mesomers, cis-trans isomers, tautomers,geometric isomers and epimers of the compound as well as the mixturethereof are included in the scope of the present invention.

The “dosage form” described herein refers to a pharmaceutical formsuitable for clinical use, including, but not limited to, pulvis,tablets, granules, capsules, solutions, emulsions, suspensions,injections (including injection solution, sterile powders for injectionand concentrated solutions for injection), sprays, aerosols, powderspray, lotions, liniments, ointments, plasters, pastes, patches, garglesor suppositories, more preferably pulvis, tablets, granules, capsules,solutions, injections, ointments, gargles or suppositories.

All technical solutions in the references cited in present applicationare included within the scope of the disclosure of the present inventionand are intended to be illustrate the present invention.

The advantages provided by the present invention include, but are notlimited to:

1. the compound and the pharmaceutically acceptable salt, ester orstereoisomer thereof have excellent inhibition activity on one or moretyrosine kinase of TRK, ALK and/or ROS1, have good pharmacokineticproperties in vivo, lasting effect and high bioavailability, and arecapable of treating and/or preventing diseases and related conditionsmediated by one or more tyrosine kinases of TRK, ALK and/or ROS1;

2. the compound and the pharmaceutically acceptable salt, ester orstereoisomer thereof have better therapeutic effect on the cancerdiseases mediated by one or more tyrosine kinases of TRK, ALK and/orROS1, and especially have better therapeutic effect on the cancerdiseases which have drug resistance to existing anti-cancer activeagents; and

3. the compound of the present invention has simple preparation processand stable quality, can be prepared with high purity, and is easy forlarge-scale industrial production.

SUMMARY

The technical solutions of the present invention will be described belowwith reference to the following examples, and the above-describedcontents of the present invention will be further described in detail,but it should not be construed that the scope of the present inventionis limited to the following examples. Any technique achieved based onthe aforementioned content of the present invention shall fall withinthe scope of the present invention.

As used herein, a carbon atom marked with an “*” in a compound structurerepresents a chiral carbon atom of a single configuration, such as an Rconfiguration or an S configuration.

In the nomenclature of compound, “2²R/S” represents that the compound isa single isomer with the configuration of the corresponding chiralcarbon being “2²R” or “2²S”

Example 1:(2²R/S,6S)-3⁵-fluoro-6-methyl-1³H-4-oxa-7-aza-1(5,3)-imidazo[4,5-b]pyridina-3(3,2)-pyridina-2(1,2)-pyrrolidinacyclooctaphan-8-one(Compound 2)

1. Preparation of6-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-3-nitropyridin-2-amine

5-fluoro-2-methoxy-3-(pyrrolidin-2-yl)pyridine (2.0 g, 10.2 mmol) and6-chloro-3-nitropyridin-2-amine (1.8 g, 10.4 mmol) were added toacetonitrile (50 mL), diisopropylethylamine (3.9 g, 30.2 mmol) wasadded, and the mixture was heated to 70° C. and reacted for 16 hrs. Thereaction mixture was concentrated by rotary evaporation, added withwater (50 mL), and filtered. The solid was washed with ethyl acetate (30mL) and dried under vacuum to give the target product (3.2 g, yield:94.1%).

LC-MS (M/e): 334.1 (M+H⁺)

2. Preparation of6-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyridine-2,3-diamine

6-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-3-nitropyridin-2-amine(1.0 g, 3.0 mmol) was dissolved in a mixture of methanol (30 mL) andtetrahydrofuran (10 mL), raney nickel (200 mg) was added, and themixture was reacted for 16 hrs at 25° C. under hydrogen atmosphere. Thereaction mixture was filtered and concentrated by rotary evaporation,and the residue was used directly in the next step.

LC-MS (M/e): 304.1 (M+H⁺)

3. Preparation of5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-3H-imidazo[4,5-b]pyridine

6-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyridine-2,3-diamine(crude product from the above step, about 3.0 mmol) was dissolved intoluene (10 mL), triethyl orthoformate (4.4 g, 29.7 mmol) andp-toluenesulfonic acid (103 mg, 0.6 mmol) were added, and the mixturewas heated to 120° C. and reacted for 4 hrs. The reaction mixture wasconcentrated by rotary evaporation, added with sodium bicarbonatesolution (50 mL), and extracted with ethyl acetate (100 mL×3). Theorganic phase was combined, concentrated by rotary evaporation andpurified by column chromatography (petroleum ether:ethyl acetate=1:1) togive the target product (0.85 g, yield: 90.4%).

LC-MS (M/e): 314.1 (M+H⁺)

4. Preparation of3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-ol

5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-3H-imidazo[4,5-b]pyridine(0.85 g, 2.72 mmol) was added to a solution of hydrogen chloride inethanol (7 mL), and the mixture was heated to 90° C. under microwave andreacted for 40 mins. The reaction mixture was concentrated by rotaryevaporation, added with triethylamine (5 mL), and concentrated by rotaryevaporation again. The reaction mixture was purified by medium-pressurepreparative chromatography (dichloromethane:methanol=15:1) to give thetarget product (0.58 g, yield: 71.4%).

LC-MS (M/e): 300.1 (M+H⁺)

5. Preparation of Tert-Butyl((2S)-1-((3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)oxy)propan-2-yl)carbamate

3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-ol(300 mg, 1.0 mmol) and tert-butyl (S)-(1-hydroxyprop-2-yl) carbamate(350 mg, 2.0 mmol) were added to tetrahydrofuran (10 mL), azobisformyldipiperidine (630 mg, 2.5 mmol) and tri-n-butylphosphine (510 mg, 2.5mmol) were added, and the mixture was reacted at 50° C. for 16 hrs. Thereaction mixture was concentrated by rotary evaporation and purified bycolumn chromatography (dichloromethane:methanol=30:1) to give the targetproduct (340 mg, yield: 74.4%).

LC-MS (M/e): 457.0 (M+H⁺)

6. Preparation of(2S)-1-((3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)oxy)propan-2-amine

Tert-butyl((2S)-1-((3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)oxy)propan-2-yl) carbamate (0.30 g, 0.66 mmol) was dissolved indichloromethane (20 mL), trifluoroacetic acid (3 mL) was added, and themixture was reacted at 25° C. for 16 hrs. The reaction mixture wasconcentrated by rotary evaporation, and the residue was used directly inthe next step.

LC-MS (M/e): 357.0 (M+H⁺)

7. Preparation of(6S)-3⁵-fluoro-6-methyl-1³H-4-oxa-7-aza-1(5,3)-imidazo[4,5-b]pyridina-3(3,2)-pyridina-2(1,2)-pyrrolidinacyclooctaphan-8-one(Compound 2-1)

A solution of(2S)-1-((3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)oxy)propan-2-amine (crude product from the above step, about 0.66 mmol) andtriethylamine (2 mL) in tetrahydrofuran (10 mL) was added dropwise to asolution of carbonyldiimidazole (160 mg, 1.0 mmol) in tetrahydrofuran(10 mL), and the mixture was reacted at 25° C. for 4 hrs. The reactionmixture was concentrated by rotary evaporation and purified bymedium-pressure preparative chromatography(dichloromethane:methanol=20:1) to give the target product (crude, 300mg).

LC-MS (M/e): 383.2 (M+H⁺)

8. Preparation of (2²RS,6S)-3⁵-fluoro-6-methyl-1³H-4-oxa-7-aza-1(5,3)-imidazo[4,5-b]pyridina-3(3,2)-pyridina-2(1,2)-pyrrolidinacyclooctaphan-8-one(Compound 2)

(6S)-3⁵-fluoro-6-methyl-1³H-4-oxa-7-aza-1(5,3)-imidazo[4,5-b]pyridina-3(3,2)-pyridina-2(1,2)-pyrrolidinacyclooctaphan-8-one (crude) was separated by high-pressurepreparative chromatography (acetonitrile:water=7:3) to give the titlecompound (50 mg, 7-9 three-step yield: 19.9%).

The title compound was prepared as a 1.12 mg/mL methanol solution andthe specific rotation of the title compound was −47.62 according to the0621 Optical Rotation Determination Method of General Chapter IV,Chinese Pharmacopoeia, 2015 Edition.

Molecular formula: C₁₉H₁₉FN₆O₂

Molecular weight: 382.4

LC-MS (M/e): 383.2 (M+H⁺)

¹H-NMR (400 MHz, CDCl₃) δ 10.01 (s, 1H), 8.38 (s, 1H), 7.88 (d, J=10.4Hz, 1H), 7.87 (s, 1H), 7.35-7.38 (m, 1H), 6.51 (d, J=9.2 Hz, 1H),5.67-5.70 (m, 1H), 5.11-5.14 (m, 1H), 4.28-4.32 (m, 1H), 4.22-4.26 (m,1H), 3.90-3.96 (m, 1H), 3.53-3.59 (m, 1H), 2.41-2.49 (m, 2H), 2.32-2.23(m, 1H), 1.92-1.95 (m, 1H), 1.59 (d, J=6.8 Hz, 3H).

Example 2:(2²R/S,6R)-3⁵-fluoro-6-methyl-1³H-4-oxa-7-aza-1(5,3)-imidazo[4,5-b]pyridina-3(3,2)-pyridina-2(1,2)-pyrrolidinacyclooctaphan-8-one(Compound 3)

1. Preparation of3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-ol

5-fluoro-3-(1-(3-(trifluoromethanesulfonyl)-3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl]pyridin-2-yltrifluoromethanesulfonate (900 mg, 1.6 mmol) was dissolved intetrahydrofuran (16 mL) and water (5 mL), lithium hydroxide monohydrate(335.2 mg, 7.99 mmol) was added, and the mixture was reacted at 25° C.for 16 hrs. The reaction mixture was adjusted to pH 6 with hydrochloricacid and extracted with ethyl acetate (30 mL). The organic phase waswashed with saturated brine (15 mL), dried over anhydrous sodiumsulfate, and concentrated to give the target product (360 mg, yield:75%).

LC-MS (M/e): 300.1 (M+H⁺)

2. Preparation of Tert-Butyl((2R)-(1-((3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)oxy)propan-2-yl)carbamate

3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-ol(360 mg, 1.2 mmol), (R)-(1-hydroxypropan-2-yl)carbamic acid tert-butylester (316.1 mg, 1.8 mmol), azobisformyl dipiperidine (455.2 mg, 1.8mmol) and tri-n-butylphosphine (365 mg, 1.8 mmol) were dissolved intetrahydrofuran (6 mL), and the mixture was reacted for 16 hrs at 50° C.under nitrogen atmosphere. The reaction mixture was concentrated andpurified by column chromatography (5% methanol/dichloromethane) to givethe target product (280 mg, yield: 51%).

LC-MS (M/e): 457.0 (M+H⁺)

3. Preparation of(2R)-1-((3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)oxy)propan-2-amine

Tert-butyl((2R)-(1-((3-(1-(3-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)oxy)propan-2-yl) carbamate (280 mg, 0.61 mmol) was dissolved indichloromethane (30 mL), trifluoroacetic acid (3 mL) was added, and themixture was reacted at 25° C. for 0.5 hr. The reaction mixture wasconcentrated by rotary evaporation, and the residue was used directly inthe next step.

LC-MS (M/e): 357.0 (M+H⁺)

4. Preparation of(6R)-3⁵-fluoro-6-methyl-1³H-4-oxa-7-aza-1(5,3)-imidazo[4,5-b]pyridina-3(3,2)-pyridina-2(1,2)-pyrrolidinacyclooctaphan-8-one(Compound 3-1)

A solution of(2R)-(1-((3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)-oxy)propan-2-amine (crude product from the above step, about 0.61 mmol) intetrahydrofuran (10 mL) and triethylamine (2 mL) were added to asolution of carbonyldiimidazole (149.2 mg, 0.92 mmol) in tetrahydrofuran(10 mL), and the mixture was reacted at 25° C. for 16 hrs. The reactionmixture was concentrated to give the target product (crude).

LC-MS (M/e): 383.2 (M+H⁺)

5. Preparation of (2²RS,6R)-3⁵-fluoro-6-methyl-1³H-4-oxa-7-aza-1(5,3)-imidazo[4,5-b]pyridina-3(3,2)-pyridina-2(1,2)-pyrrolidinacyclooctaphan-8-one(Compound 3)

(6R)-3⁵-fluoro-6-methyl-1³H-4-oxa-7-aza-1(5,3)-imidazo[4,5-b]pyridina-3(3,2)-pyridina-2(1,2)-pyrrolidinacyclooctaphan-8-one (crude) was separated by high-pressurereversed-phase column chromatography (acetonitrile:water=7:3) to givethe title compound (12 mg, 3-5 three-step yield: 5.1%).

Molecular formula: C₁₉H₁₉FN₆O₂

Molecular weight: 382.4

LC-MS (M/e): 383.0 (M+H⁺)

¹H-NMR (400 MHz, CDCl₃) δ 10.50 (d, J=7.6 Hz, 1H), 8.35 (s, 1H),7.89-7.84 (m, 2H), 7.34-7.26 (m, 1H), 6.50 (d, J=8.8 Hz, 1H), 5.63-5.60(m, 1H), 5.16-5.12 (m, 1H), 4.45-4.35 (m, 1H), 4.18-4.13 (m, 1H),3.95-3.92 (m, 1H), 3.62-3.59 (m, 1H), 2.54-2.42 (m, 2H), 2.26-2.23 (m,1H), 1.99-1.96 (m, 1H), 1.56 (d, J=8.8 Hz, 3H).

Example 3:(2²R/S,6R)-3⁵-fluoro-6-methyl-1³H-7-aza-1(5,3)-imidazo[4,5-b]pyridina-3(3,2)-pyridina-2(1,2)-pyrrolidinacyclooctaphan-8-one(Compound 11)

1. Preparation of Benzyl(R)-but-3-yn-2-yl((2-nitrophenyl)sulfonyl)carbamate

Benzyl ((2-nitrophenyl)sulfonyl)carbamate (5.0 g, 14.9 mmol) and(S)-but-3-yn-2-ol (1.0 g, 14.3 mmol) were dissolved in tetrahydrofuran(60 mL), and triphenylphosphine (4.0 g, 15.2 mmol) was added. Diethylazodicarboxylate (2.7 g, 15.5 mmol) was added under nitrogen atmosphere,and the mixture was reacted at 20° C. for 16 hrs. The reaction mixturewas concentrated and purified by column chromatography (petroleumether:ethyl acetate=2:1) to give the target product (3.16 g, yield:56.9%).

LC-MS (M/e): 389.0 (M+H⁺)

2. Preparation of Benzyl (R)-but-3-yn-2-yl Carbamate

Benzyl (R)-but-3-yn-2-yl((2-nitrophenyl)sulfonyl)carbamate (3.16 g, 8.14mmol) was dissolved in N,N-dimethylformamide (30 mL), and lithiumhydroxide (2.4 g, 57.1 mmol) and thioglycolic acid (2.6 g, 28.3 mmol)were added, and the mixture was reacted at 20° C. for 16 hrs. Thereaction mixture was added with water (100 mL) and ethyl acetate (100mL) and separated, and the aqueous phase was extracted with ethylacetate (100 mL×3). The organic phase was combined, concentrated andpurified by column chromatography (petroleum ether:ethyl acetate=10:1)to give the target product (890 mg, yield: 53.9%).

LC-MS (M/e): 204.1 (M+H⁺)

3. Preparation of6-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-3-nitropyridin-2-amine

5-fluoro-2-methoxy-3-(pyrrolidin-2-yl)pyridine (2.0 g, 10.2 mmol) and6-chloro-3-nitropyridin-2-amine (1.8 g, 10.4 mmol) were added toacetonitrile (50 mL), diisopropylethylamine (3.9 g, 30.2 mmol) wasadded, and the mixture was heated to 70° C. and reacted for 16 hrs. Thereaction mixture was concentrated by rotary evaporation, added withwater (50 mL) and filtered; the solid was washed with ethyl acetate (30mL) and dried under vacuum to give the target product (3.2 g, yield:94.1%).

LC-MS (M/e): 334.1 (M+H⁺)

4. Preparation of6-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyridine-2,3-diamine

6-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-3-nitropyridin-2-amine(1.0 g, 3.0 mmol) was dissolved in a mixture of methanol (30 mL) andtetrahydrofuran (10 mL), raney nickel (200 mg) was added, and themixture was reacted for 16 hrs at 25° C. under hydrogen atmosphere. Thereaction mixture was filtered and concentrated by rotary evaporation,and the residue was used directly in the next step.

LC-MS (M/e): 304.1 (M+H⁺)

5. Preparation of5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-3H-imidazo[4,5-b]pyridine

6-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyridine-2,3-diamine(crude product from the above step, about 3.0 mmol) was dissolved intoluene (10 mL), triethyl orthoformate (4.4 g, 29.7 mmol) andp-toluenesulfonic acid (103 mg, 0.6 mmol) were added, and the mixturewas heated to 120° C. and reacted for 4 hrs. The reaction mixture wasconcentrated by rotary evaporation, added with sodium bicarbonatesolution (50 mL), and extracted with ethyl acetate (100 mL×3). Theorganic phase was combined, concentrated by rotary evaporation andpurified by column chromatography (petroleum ether:ethyl acetate=1:1) togive the target product (0.85 g, yield: 90.4%).

LC-MS (M/e): 314.1 (M+H⁺)

6. Preparation of3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl-5-fluoropyridin-2-ol

5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-3H-imidazo[4,5-b]pyridine(0.85 g, 2.72 mmol) was added to a solution of hydrogen chloride inethanol (7 mL), and the mixture was heated to 90° C. under microwave andreacted for 40 mins. The reaction mixture was concentrated by rotaryevaporation, added with triethylamine (5 mL), and concentrated by rotaryevaporation again. The reaction mixture was purified by medium-pressurepreparative chromatography (dichloromethane:methanol=15:1) to give thetarget product (0.58 g, yield: 71.4%).

LC-MS (M/e): 300.1 (M+H⁺)

7. Preparation of5-fluoro-3-(1-(3-((trifluoromethyl)sulfonyl)-3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)pyridin-2-ylTrifluoromethanesulfonate

3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-ol(0.58 g, 1.94 mmol) was dissolved in dichloromethane (20 mL),1,1,1-trifluoro-N-phenyl-N-((trifluoromethyl)sulfonyl)methanesulfonamide (2.1 g, 5.88 mmol) and triethylamine (784 mg, 7.76mmol) were added, and the mixture was reacted at 20° C. for 16 hrs. Thereaction mixture was concentrated by rotary evaporation and purified bycolumn chromatography (petroleum ether:ethyl acetate=2:1) to give thetarget product (0.73 g, yield: 67.0%).

LC-MS (M/e): 564.0 (M+H⁺)

8. Preparation of Benzyl((2R)-4-(5-fluoro-3-(1-(3-((trifluoromethyl)sulfonyl)-3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)pyridin-2-yl)-but-3-yn-2-yl)carbamate

5-fluoro-3-(1-(3-((trifluoromethyl)sulfonyl)-3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)pyridin-2-yltrifluoromethanesulfonate (0.73 g, 1.3 mmol), cuprous iodide (50 mg,0.26 mmol), bis(triphenylphosphine)palladium(II) dichloride (91 mg, 0.13mmol) and N,N-diisopropylethylamine (503 mg, 3.9 mmol) were dissolved intetrahydrofuran (50 mL), and the mixture was reacted for 0.5 hr at 50°C. under nitrogen atmosphere. A solution of benzyl (R)-but-3-yn-2-ylcarbamate (300 mg, 1.5 mmol) in tetrahydrofuran was added slowly, andthe mixture was reacted at 50° C. for 4 hrs. The reaction mixture wasconcentrated by rotary evaporation and purified by column chromatography(petroleum ether:ethyl acetate=1:1) to give the target product (0.53 g,yield: 66.3%).

LC-MS (M/e): 617.2 (M+H⁺)

9. Preparation of Benzyl((2R)-4-(3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)-but-3-yn-2-yl)carbamate

Benzyl((2R)-4-(5-fluoro-3-(1-(3-((trifluoromethyl)sulfonyl)-3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)pyridin-2-yl)-but-3-yn-2-yl)carbamate(0.53 g, 0.86 mmol) was dissolved in tetrahydrofuran (20 mL) and water(20 mL), lithium hydroxide (180 mg, 4.3 mmol) was added, and the mixturewas reacted at 20° C. for 1 hr. The reaction mixture was added withethyl acetate (20 mL) and separated, and the aqueous phase was extractedwith ethyl acetate (20 mL×2). The organic phase was combined,concentrated and purified by column chromatography(dichloromethane:methanol=20:1) to give the target product (0.4 g,yield: 96.2%).

LC-MS (M/e): 485.2 (M+H⁺)

10. Preparation of(2R)-4-(3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)butan-2-amine

Benzyl((2R)-4-(3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)-but-3-yn-2-yl)carbamate(0.4 g, 0.83 mmol) was dissolved in methanol (50 mL), Palladium onCarbon (200 mg) was added, and the mixture was reacted for 16 hrs at 20°C. under hydrogen atmosphere. The reaction mixture was filtered, thefiltrate was concentrated to give the target product (crude) which wasused directly in the next step without purification.

LC-MS (M/e): 355.2 (M+H⁺)

11. Preparation of(6R)-3⁵-fluoro-6-methyl-1³H-7-aza-1(5,3)-imidazo[4,5-b]pyridina-3(3,2)-pyridina-2(1,2)-pyrrolidinacyclooctaphan-8-one(Compound 11-1)

N,N′-carbonyldiimidazole (250 mg, 1.54 mmol) was dissolved intetrahydrofuran (20 mL), and then added to a solution of(2R)-4-(3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)butan-2-amine (crude product from the above step) in tetrahydrofuran (10mL), and the mixture was reacted at 25° C. for 2 hrs. The reactionmixture was concentrated and purified by column chromatography(dichloromethane:methanol=20:1) to give the target product (crude, 40mg).

12. Preparation of(2²R/S,6R)-3⁵-fluoro-6-methyl-1³H-7-aza-1(5,3)-imidazo[4,5-b]pyridina-3(3,2)-pyridina-2(1,2)-pyrrolidinacyclooctaphan-8-one (Compound 11)

(6R)-3⁵-fluoro-6-methyl-1³H-7-aza-1(5,3)-imidazo[4,5-b]pyridina-3(3,2)-pyridina-2(1,2)-pyrrolidinacyclooctaphan-8-one (crude) was separated by high-pressure preparativechromatography (acetonitrile:water=8:2) to give the title compound (3mg, 10-12 three-step yield: 0.95%).

Molecular formula: C₂₀H₂₁FN₆O

Molecular weight: 380.4

LC-MS (M/e): 381.1 (M+H⁺)

¹H-NMR (400 MHz, CDCl₃) δ 9.46 (d, J=8.0 Hz, 1H), 8.33 (s, 1H), 8.30 (d,J=2.8 Hz, 1H), 7.88 (d, J=8.8 Hz, 1H), 7.14 (dd, J=2.8 Hz, 9.6 Hz, 1H),6.53 (d, J=8.8 Hz, 1H), 5.42 (t, J=6.0 Hz, 1H), 4.35-4.37 (m, 1H),3.91-3.96 (m, 1H), 3.68-3.71 (m, 1H), 3.39-3.43 (m, 1H), 2.87-2.92 (m,2H), 2.51-2.54 (m, 1H), 2.34-2.38 (m, 1H), 2.18-2.25 (m, 2H), 2.00-2.02(m, 1H), 1.83-1.88 (m, 1H), 1.39 (d, J=6.8 Hz, 3H).

Example 4:(3R,7S)-4⁵-fluoro-3,7-dimethyl-1³H-5-oxa-2,8-diaza-1(5,3)-imidazo[4,5-b]pyridina-4(3,2)-pyridinacyclononphan-9-one(Compound 13)

1. Preparation of3-(1-((3H-imidazo[4,5-b]pyridin-5-ylamino)ethyl)-5-fluoropyridin-2-ol

Tert-butyl5-((1-(5-fluoro-2-methoxypyridin-3-yl)ethyl)amino)-3H-imidazo[4,5-b]pyridine-3-carboxylate(400 mg, 104 mmol) was dissolved in a solution of hydrogen chloride inethanol (6 mL), and the mixture was reacted at 85° C. for 16 hrs. Thereaction mixture was concentrated by rotary evaporation, adjusted to pH8 with triethylamine, and purified by reversed-phase columnchromatography (0-20% acetonitrile/water) to give the target product(100 mg, yield: 35%).

LC-MS (M/e): 274.0 (M+H⁺)

2. Preparation of Tert-Butyl((2S)-1-((3-(1-((3H-imidazo[4,5-b]pyridin-5-yl)amino)ethyl)-5-fluoropyridin-2-yl)oxy))propan-2-yl)carbamate

3-(1-((3H-imidazo[4,5-b]pyridin-5-yl amino)ethyl)-5-fluoropyridin-2-ol(100 mg, 0.37 mmol), tert-butyl (S)-(1-hydroxypropan-2-yl)carbamate(96.2 mg, 0.55 mmol), azobisformyl dipiperidine (138.5 mg, 0.55 mmol)and tri-n-butylphosphine (111.1 mg, 0.55 mmol) were dissolved intetrahydrofuran (2 mL), and the mixture was reacted for 16 hrs at 50° C.under nitrogen atmosphere. The reaction mixture was concentrated andpurified by column chromatography (5% methanol/dichloromethane) to givethe target product (40 mg, yield: 25%).

LC-MS (M/e): 431.0 (M+H⁺)

3. Preparation ofN-(1-(2-((S)-2-aminopropoxy)-5-fluoropyridin-3-yl)ethyl)-3H-imidazo[4,5-b]pyridin-5-amine

Tert-butyl((2S)-1-((3-(1-((3H-imidazo[4,5-b]pyridin-5-yl)amino)ethyl)-5-fluoropyridin-2-yl)oxy))propan-2-yl)carbamate (40 mg, 0.093 mmol) was dissolved indichloromethane (2 mL), trifluoroacetic acid (1 mL) was added, and themixture was reacted for 0.5 hr. The reaction mixture was concentratedand the crude product was used directly in the next step.

LC-MS (M/e): 331.0 (M+H⁺)

4. Preparation of(7S)-4⁵-fluoro-3,7-dimethyl-1³H-5-oxa-2,8-diaza-1(5,3)-imidazo[4,5-b]pyridina-4(3,2)-pyridinacyclononphan-9-one(Compound 13-1)

A solution ofN-(1-(2-((S)-2-aminopropoxy)-5-fluoropyridin-3-yl)ethyl)-3H-imidazo[4,5-b]pyridin-5-amine(crude product from the above step) in tetrahydrofuran (1 mL) andtriethylamine (0.2 mL) were added to a solution of carbonyldiimidazole(22.6 mg, 0.14 mmol) in tetrahydrofuran (1 mL), and the mixture wasreacted at 25° C. for 16 hrs. The reaction mixture was concentrated togive the target product (crude).

LC-MS (M/e): 357.1 (M+H⁺)

5. Preparation of(3R,7S)-4⁵-fluoro-3,7-dimethyl-1³H-5-oxa-2,8-diaza-1(5,3)-imidazo[4,5-b]pyridina-4(3,2)-pyridinacyclononphan-9-one(Compound 13)

(7S)-4⁵-fluoro-3,7-dimethyl-1³H-5-oxa-2,8-diaza-1(5,3)-imidazo[4,5-b]pyridina-4(3,2)-pyridinacyclononphan-9-one was separated by high-pressure reversed-phase columnchromatography (acetonitrile/water=70%) to give the title compound (3mg, 3-5 three-step yield: 9%).

The title compound was prepared as a 1.12 mg/mL methanol solution andthe specific rotation of the title compound was +175.3 according to the0621 Optical Rotation Determination Method of General Chapter IV,Chinese Pharmacopoeia, 2015 Edition.

Molecular formula: C₁₇H₁₇FN₆O₂

Molecular weight: 356.4

LC-MS (M/e): 357.1 (M+H⁺)

¹H-NMR (400 MHz, CDCl₃) δ 10.28 (s, 1H), 8.37 (s, 1H), 7.88 (d, J=2.8,1H), 7.79 (d, J=8.8, 1H), 7.48-7.44 (m, 1H), 6.43 (d, J=8.8, 1H),5.56-5.53 (m, 1H), 5.06-5.03 (m, 1H), 4.92-4.89 (m, 1H), 4.31-4.25 (m,2H), 1.63 (d, J=6.4, 3H), 1.53 (d, J=6.8, 3H).

Example 5:(2²R,6R)-3⁵-fluoro-6-methyl-1³H-4-oxa-7-aza-1(5,3)-imidazo[4,5-b]pyridina-3(3,2)-pyridina-2(1,2)-pyrrolidinacyclooctaphan-8-one(Compound 3′)

1. Preparation of(R)-6-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-3-nitropyridin-2amine

(R)-5-fluoro-2-methoxy-3-(pyrrolidin-2-yl)pyridine (1.0 g, 5.1 mmol) and6-chloro-3-nitropyridin-2-amine (886 mg, 5.1 mmol) were added toacetonitrile (30 mL), diisopropylethylamine (1.98 g, 15.3 mmol) wasadded, and the mixture was heated to 70° C. and reacted for 16 hrs. Thereaction mixture was concentrated by rotary evaporation, added withwater (30 mL), and filtered. The solid was washed with ethyl acetate (30mL) and dried under vacuum to give the target product (1.65 g, yield:97.0%).

LC-MS (M/e): 334.1 (M+H⁺)

2. Preparation of(R)-6-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyridine-2,3-diamine

(R)-6-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-3-nitropyridin-2-amine(1.65 g, 4.94 mmol) was dissolved in methanol (20 mL), raney nickel (1g) was added, and the mixture was reacted for 16 hrs at 20° C. underhydrogen atmosphere. The reaction mixture was filtered and concentratedby rotary evaporation, and the residue was used directly in the nextstep.

LC-MS (M/e): 304.1 (M+H⁺)

3. Preparation of(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-3H-imidazo[4,5-b]pyridine

(R)-6-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyridine-2,3-diamine(crude product from the above step, about 3.0 mmol) was dissolved intoluene (30 mL), trimethyl orthoformate (5.2 g, 49 mmol) andp-toluenesulfonic acid (170 mg, 0.99 mmol) were added, and the mixturewas heated to 110° C. and reacted for 5 hrs. The reaction mixture wasconcentrated by rotary evaporation, added with a sodium bicarbonatesolution (100 mL) and ethyl acetate (100 mL), and separated, and theaqueous phase was extracted with ethyl acetate (50 mL×3). The organicphase was combined, concentrated by rotary evaporation and purified bycolumn chromatography (ethyl acetate) to give the target product (1.5 g,yield: 96.8%).

LC-MS (M/e): 314.1 (M+H⁺)

4. Preparation of(R)-3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-ol

(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-3H-imidazo[4,5-b]pyridine(1.5 g, 4.79 mmol) was added to a solution of hydrogen chloride inethanol (30 mL), and the mixture was heated to 90° C. and reacted for 16hrs. The reaction mixture was concentrated by rotary evaporation,adjusted pH to alkaline with the addition of triethylamine (5 mL),concentrated by rotary evaporation again, and purified by columnchromatography (dichloromethane:methanol=10:1) to give the targetproduct (1.4 g, yield: 97.9%).

LC-MS (M/e): 300.1 (M+H⁺)

5. Preparation of Tert-Butyl((R)-1-((3-((R)-1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)oxy)propan-2-yl)carbamate

(R)-3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-ol(700 mg, 2.34 mmol), tert-butyl (R)-(1-hydroxypropan-2-yl)carbamate (410mg, 2.34 mmol) and tri-n-butylphosphine (945 mg, 4.68 mmol) weredissolved in tetrahydrofuran (10 mL), and azobisformyl dipiperidine(1.18 g, 4.68 mmol) was added under nitrogen atmosphere, and the mixturewas reacted at 30° C. for 2 hrs. The reaction mixture was concentrated,and purified by C18 column chromatography to give the target product(500 mg, yield: 46.7%).

LC-MS (M/e): 457.0 (M+H⁺)

6. Preparation of(R)-1-((3-((R)-1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)oxy)propan-2-amine

Tert-butyl((R)-1-((3-((R)-1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)oxy)propan-2-yl) carbamate (500 mg, 1.1 mmol) was dissolved indichloromethane (20 mL), trifluoroacetic acid (20 mL) was added, and themixture was reacted at 20° C. for 16 hrs. The reaction solution wasconcentrated, adjusted pH to alkaline with triethylamine, concentratedby rotary evaporation, and purified by C18 column chromatography to givethe target product (crude, 400 mg).

LC-MS (M/e): 357.0 (M+H⁺)

7. Preparation of(2²R,6R)-3⁵-fluoro-6-methyl-1³H-4-oxa-7-aza-1(5,3)-imidazo[4,5-b]pyridina-3(3,2)-pyridina-2(1,2)-pyrrolidinacyclooctaphan-8-one(Compound 3)

(R)-1-((3-((R)-1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)oxy)propan-2-amine(crude product from the above step, 400 mg, about 1.1 mmol) wasdissolved in xylene (50 mL), carbonyldiimidazole (535 mg, 3.3 mmol) wasadded, and the mixture was warmed to 130° C. and reacted for 2 hrs. Thereaction mixture was concentrated, purified by column chromatography(petroleum ether:ethyl acetate=1:1) to give the target product (crude,300 mg), and further separated by HPLC high-pressure reversed-phasepreparative chromatography (acetonitrile:water=10%-70%) to give thetitle compound (100 mg, yield: 23.8%).

Molecular formula: C₁₉H₁₉FN₆O₂

Molecular weight: 382.4

LC-MS (M/e): 383.2 (M+H⁺)

¹H-NMR (400 MHz, CDCl₃) δ 10.50 (m, 1H), 8.36 (s, 1H), 7.86-7.90 (m,2H), 7.31-7.35 (m, 1H), 6.51 (d, J=8.8 Hz, 1H), 5.63-5.60 (m, 1H),5.13-5.17 (m, 1H), 4.39-4.42 (m, 1H), 4.15-4.19 (m, 1H), 3.92-3.98 (m,1H), 3.65-3.60 (m, 1H), 2.50-2.59 (m, 2H), 2.40-2.50 (m, 1H), 1.95-2.01(m, 1H), 1.57 (d, J=6.8 Hz, 3H).

The compound 3 and the compound 3′ were the same compound, which wasconfirmed by methods including hydrogen spectrum, mass spectrum, HPLC,specific rotation and the like.

Example 6:(2²R,6S)-3⁵-fluoro-6-methyl-1³H-4-oxa-7-aza-1(5,3)-imidazo[4,5-b]pyridina-3(3,2)-pyridina-2(1,2)-pyrrolidinacyclooctaphan-8-one(Compound 2′)

The preparation thereof was the same as in Example 5 except thattert-butyl (S)-(1-hydroxypropan-2-yl)carbamate was used in place oftert-butyl (R)-(1-hydroxypropan-2-yl)carbamate in the step 5.

The compound 2 and the compound 2′ were the same compound, which wasconfirmed by methods including hydrogen spectrum, mass spectrum, HPLC,specific rotation and the like.

Example 7:(2²R,6R)-3⁵-fluoro-6-methyl-1³H-7-aza-1(5,3)-imidazo[4,5-b]pyridina-3(3,2)-pyridina-2(1,2)-pyrrolidinacyclooctaphan-8-one_(Compound11′)

1. Preparation of(R)-6-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-3-nitropyridin-2-amine

(R)-5-fluoro-2-methoxy-3-(pyrrolidin-2-yl)pyridine (2.0 g, 10.2 mmol)and 6-chloro-3-nitropyridin-2-amine (1.8 g, 10.4 mmol) were added toacetonitrile (50 mL), diisopropylethylamine (3.9 g, 30.2 mmol) wasadded, and the mixture was heated to 70° C. for 16 hrs. The reactionmixture was concentrated by rotary evaporation, added with water (50mL), and filtered. The solid was washed with ethyl acetate (30 mL) anddried under vacuum to give the target product (3.27 g, yield: 96.2%).

LC-MS (M/e): 334.1 (M+H⁺)

2. Preparation of(R)-6-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyridine-2,3-diamine

(R)-6-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-3-nitropyridin-2-amine(3.27 g, 9.8 mmol) was dissolved in methanol (50 mL), raney nickel (2 g)was added, and the mixture was reacted for 3 hrs at 20° C. underhydrogen atmosphere. The reaction mixture was filtered and concentratedby rotary evaporation, and the residue was used directly in the nextstep.

LC-MS (M/e): 304.1 (M+H⁺)

3. Preparation of(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-3H-imidazo[4,5-b]pyridine

(R)-6-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)pyridine-2,3-diamine(crude product from the above step, about 9.8 mmol) was dissolved intoluene (100 mL), trimethyl orthoformate (10.4 g, 98 mmol) andp-toluenesulfonic acid (170 mg, 0.99 mmol) were added, and the mixturewas heated to 110° C. and reacted for 16 hrs. The reaction mixture wasconcentrated by rotary evaporation, added with sodium bicarbonatesolution (100 mL), and extracted with ethyl acetate (100 mL×3). Theorganic phase was combined, concentrated by rotary evaporation andpurified by column chromatography (petroleum ether:ethyl acetate=1:1) togive the target product (3.05 g, yield: 99.0%).

LC-MS (M/e): 314.1 (M+H⁺)

4. Preparation of(R)-3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-ol

(R)-5-(2-(5-fluoro-2-methoxypyridin-3-yl)pyrrolidin-1-yl)-3H-imidazo[4,5-b]pyridine(3.05 g, 9.7 mmol) was added to a solution of hydrogen chloride inethanol (100 mL), and the mixture was heated to 80° C. and reacted for16 hrs. The reaction mixture was concentrated by rotary evaporation,added with triethylamine (5 mL), and concentrated by rotary evaporationagain. The reaction mixture was purified by column chromatography(dichloromethane:methanol=10:1) to give the target product (2.8 g,yield: 96.2%).

LC-MS (M/e): 300.1 (M+H⁺)

5. Preparation of(R)-5-fluoro-3-(1-(3-((trifluoromethyl)sulfonyl)-3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)pyridin-2-yl Trifluoromethanesulfonate

(R)-3-(1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-ol(2.8 g, 9.36 mmol) was dissolved in dichloromethane (100 mL),1,1,1-trifluoro-N-phenyl-N-((trifluoromethyl)sulfonyl)methanesulfonamide (10 g, 28 mmol) and triethylamine (4.7 g, 46.5 mmol)were added, and the mixture was reacted at 20° C. for 16 hrs. Thereaction mixture was concentrated by rotary evaporation and purified bycolumn chromatography (petroleum ether:ethyl acetate=5:1) to give thetarget product (4.46 g, yield: 84.6%).

LC-MS (M/e): 564.0 (M+H⁺)

6. Preparation of Benzyl((R)-4-(5-fluoro-3-((R)-1-(3-((trifluoromethyl)sulfonyl)-3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)pyridin-2-yl)-but-3-yn-2-yl)carbamate

(R)-5-fluoro-3-(1-(3-((trifluoromethyl)sulfonyl)-3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)pyridin-2-yltrifluoromethanesulfonate (4.46 g, 7.92 mmol), cuprous iodide (300 mg,1.48 mmol), bis(triphenylphosphine)palladium dichloride (560 mg, 0.8mmol) and DIPEA (3.0 g, 23.2 mmol) were dissolved in tetrahydrofuran(100 mL), and the mixture was reacted for 0.5 hr at 50° C. undernitrogen atmosphere. A solution of benzyl (R)-but-3-yn-2-yl carbamate(1.6 g, 7.9 mmol) in tetrahydrofuran was added slowly, and the mixturewas reacted at 50° C. for 5 hrs. The reaction mixture was concentratedby rotary evaporation and purified by column chromatography (petroleumether:ethyl acetate=2:1) to give the target product (3.7 g, yield:75.8%).

LC-MS (M/e): 617.2 (M+H⁺)

7. Preparation of Benzyl((R)-4-(3-((R)-1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)-but-3-yn-2-yl)carbamate

Benzyl((R)-4-(5-fluoro-3-((R)-1-(3-((trifluoromethyl)sulfonyl)-3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)pyridin-2-yl)-but-3-yn-2-yl)carbamate (3.7 g, 6.0 mmol)was dissolved in tetrahydrofuran (40 mL) and water (10 mL), lithiumhydroxide (1.26 g, 30 mmol) was added, and the mixture was reacted at20° C. for 2 hrs. The reaction mixture was added with ethyl acetate (100mL) and water (50 mL) and separated, and the aqueous phase was extractedwith ethyl acetate (50 mL×2). The organic phase was combined,concentrated and purified by column chromatography(dichloromethane:methanol=20:1) to give the target product (2.6 g,yield: 89.6%).

LC-MS (M/e): 485.2 (M+H⁺)

8. Preparation of(R)-4-(3-((R)-1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)-butan-2-amine

Benzyl((R)-4-(3-((R)-1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)-but-3-yn-2-yl)carbamate(1 g, 2.1 mmol) was dissolved in methanol (50 mL), Palladium on Carbon(500 mg) was added, and the mixture was reacted for 16 hrs at 20° C.under hydrogen atmosphere. The reaction mixture was filtered, thefiltrate was concentrated to give the target product (crude, 700 mg)which was used directly in the next step without purification.

LC-MS (M/e): 335.2 (M+H⁺)

9. Preparation of(2²R,6R)-3⁵-fluoro-6-methyl-1³H-7-aza-1(5,3)-imidazo[4,5-b]pyridina-3(3,2)-pyridina-2(1,2)-pyrrolidinacyclooctaphan-8-one(Compound 11)

N,N-carbonyldiimidazole (412 mg, 2.54 mmol) was dissolved in xylene (25mL), and then a solution of(R)-4-(3-((R)-1-(3H-imidazo[4,5-b]pyridin-5-yl)pyrrolidin-2-yl)-5-fluoropyridin-2-yl)-butan-2-amine(crude, 300 mg, about 0.85 mmol) in xylene (25 mL) was added, and themixture was reacted at 20° C. for 2 hrs, warmed to 130° C. and reactedfor 5 hrs. The reaction mixture was concentrated, purified by columnchromatography (petroleum ether:ethyl acetate=1:1) to give the targetproduct (crude), and separated by HPLC high-pressure preparativechromatography (acetonitrile:water=10%-80%) to give the title compound(70 mg, yield: 21.7%).

Molecular formula: C₂₀H₂₁FN₆O

Molecular weight: 380.4

LC-MS (M/e): 381.1 (M+H⁺)

¹H-NMR (400 MHz, CDCl₃) δ 9.48 (d, J=8.4 Hz, 1H), 8.35 (s, 1H), 8.31 (d,J=2.4 Hz, 1H), 7.89 (d, J=8.8 Hz, 1H), 7.15 (dd, J₁=2.4 Hz, J₂=9.6 Hz,1H), 6.55 (d, J=8.8 Hz, 1H), 5.43 (t, J=6.4 Hz, 1H), 4.34-4.37 (m, 1H),3.93-3.99 (m, 1H), 3.68-3.74 (m, 1H), 3.40-3.47 (m, 1H), 2.86-2.98 (m,2H), 2.50-2.57 (m, 1H), 2.36-2.41 (m, 1H), 2.17-2.27 (m, 2H), 1.87-1.98(m, 1H), 1.82-1.89 (m, 1H), 1.40 (d, J=6.8 Hz, 3H).

The compound 11 and the compound 11′ were the same compound, which wasconfirmed by methods including hydrogen spectrum, mass spectrum, HPLC,specific rotation and the like.

Activity Assay

Exemplary activity assay for some compounds of the present invention areprovided hereinafter to show the beneficial activity and beneficialtechnical effect of the compound of the present invention. However, itshould be understood that the following experimental schemes are onlyillustrations for the present invention rather than limit the scope ofthe present invention.

Experimental Example 1: In Vitro Cytological Inhibitory Activity ofCompounds of the Present Invention Experimental Materials

The test compounds were compounds in the examples above.

The cell lines used in the experiment have the following meanings:

Ba/F3 LMNA-NTRK1-G595R cell line:

a stable expression cell line of Ba/F3 cells transfected with LMNA-NTRK1G595R;

Ba/F3 ETV6-NTRK3-G623R cell line:

a stable expression cell line of Ba/F3 cells transfected withETV6-NTRK3-G623R.

Experimental Method (CelltiterGlo Assay)

1. Cell Culture and Inoculation

All cells were suspension cells and the medium was RPMI-1640+10% FBS,and cells were tested in a logarithmic growth phase.

Cells in the logarithmic growth phase were collected and counted using aplatelet counter. The cell viability was detected by the trypan blueexclusion method, and the cell viability was kept over 90%. The cellconcentration was adjusted to be in a proper range; 90 μL of the cellsuspension was added to a 96-well plate.

TABLE 1 Number of cell inoculation Cell lines Number of cells inoculatedBa/F3 LMNA-NTRK1-G595R 3000 cells/well Ba/F3 ETV6-NTRK3-G623R 3000cells/well

2. Preparation of Test Compounds

2.1 Preparation of DMSO Stock Solutions of Test Compounds, See Table 2for Concentrations Thereof.

TABLE 2 Stock solution concentration (mM) of test compounds CompoundsStock solution concentration Compound 2 10 mM Compound 3 10 mM Compound11 10 mM Compound 13 10 mM

2.2 Preparation of Working Stock Solutions of Test Compounds

The stock solutions of test compounds were diluted from 10 mM to 1 mMwith DMSO and then serially diluted in a 3-fold gradient for 9concentrations. The 2 μL DMSO-gradient diluted compound solutions wereadded to 98 μL culture solutions to give working stock solutions of testcompounds (compound concentration was 10-fold higher than the finalconcentration, DMSO concentration was 1%, maximum concentration was 10μM).

Maximum concentration was in DMSO solvent control, blank wells were onlyadded with medium without cells inoculated.

2.3 Compound Treatment

10 μL of compound working stock solution was added to each well in the96-well plate inoculated with cells (10-fold dilution, final DMSOconcentration was 0.1%).

The final concentrations of test compounds were: 1000 nM, 333.33 nM,111.11 nM, 37.04 nM, 12.35 nM, 4.12 nM, 1.37 nM, 0.46 nM and 0.15 nM.

The cells were cultured in a cell culture incubator at 5% CO₂ for 72hrs.

3. Detection

The CTG reagent was thawed and the cell plate was equilibrated to roomtemperature for 30 mins, 60 μL of reagent (Celltiter Glo assay kit) wasadded to each well, shaking was performed for 2 mins with a shaker (inthe absence of light), and the plate was incubated at room temperaturefor 10 mins (in the absence of light). The light signal values were readon the multi-functional microplate reader.

4. Data Processing

1) Inhibition (%)=(DMSO solvent control well reading−test compound wellreading)/(DMSO solvent control well reading−blank well reading)×100%;

2) a curve was plotted with GraphPad Prism 5.0 and IC₅₀ was obtained.

Experimental Result and Conclusion

TABLE 3 In vitro cytological activity (IC₅₀, nM) of compounds of thepresent invention Ba/F3 LMNA-NTRK1- Ba/F3 ETV6-NTRK3- Test compoundsG595R(nM) G623R(nM) Compound 2 0.6 1 Compound 3 0.6 1 Compound 11 2 1.9Compound 13 17.3 18.9

It can be seen from Table 3 that the compounds of the present inventioncan effectively inhibit the proliferation of cells such as Ba/F3LMNA-NTRK1-G595R and Ba/F3 ETV6-NTRK3-G623R, indicating that thecompounds of the present invention have the potential of clinicalapplication in treating drug-resistant cancerous diseases caused by NTRKgene mutation.

Experimental Example 2: In Vitro Cytological Inhibitory Activity ofCompounds of the Present Invention Experimental Materials

The test compounds were compounds in the examples above.

The cell lines used in the experiment have the following meanings:

Ba/F3 SLC34A2/ROS1-G2032R cell line:

a stable expression cell line of Ba/F3 cells transfected withSLC34A2/ROS1-G2032R.

Experimental Method (Celltiter-Glo Assay)

1. Cell Culture and Inoculation

All cells were suspension cells and the medium was RPMI-1640+10% FBS,and cells were tested in a logarithmic growth phase.

Cells in the logarithmic growth phase were collected and counted using aplatelet counter. The cell viability was detected by the trypan blueexclusion method, and the cell viability was kept over 90%. The cellconcentration was adjusted to be in a proper range; 90 μL of the cellsuspension was added to a 96-well plate.

TABLE 4 Number of cell inoculation Cell lines Number of cells inoculatedBa/F3 SLC34A2/ROS1-G2032R 3000 cells/well

2. Preparation of Test Compounds

2.1 Preparation of DMSO Stock Solutions of Test Compounds, See Table 5for Concentrations Thereof.

TABLE 5 Stock solution concentration (mM) of test compounds CompoundsStock solution concentration Compound 3 10 mM Compound 11 10 mM Compound13 10 mM

2.2 Preparation of Working Stock Solutions of Test Compounds

The stock solutions of test compounds were diluted from 10 mM to 1 mMwith DMSO and then serially diluted in a 3.16-fold gradient for 9concentrations. The 2 μL DMSO-gradient diluted compound solutions wereadded to 98 μL culture solutions to give working stock solutions of testcompounds (compound concentration was 10-fold higher than the finalconcentration, DMSO concentration was 1%, maximum concentration was 10μM).

Maximum concentration was in DMSO solvent control, blank wells were onlyadded with medium without cells inoculated.

2.3 Compound Treatment

10 μL of compound working stock solution was added to each well in the96-well plate inoculated with cells (10-fold dilution, final DMSOconcentration was 0.1%).

The final concentrations of test compounds were: 1000 nM, 316 nM, 100nM, 31.6 nM, 10 nM, 3.16 nM, 1 nM, 0.316 nM and 0.1 nM.

The cells were cultured in a cell culture incubator at 5% CO₂ for 72hrs.

3. Detection

The CTG reagent was thawed and the cell plate was equilibrated to roomtemperature for 30 mins, 100 μL of reagent (Celititer Glo assay kit) wasadded to each well, shaking was performed for 5 mins with a shaker (inthe absence of light), and the plate was incubated at room temperaturefor 20 mins (in the absence of light). The light signal values were readon the multi-functional microplate reader.

4. Data Processing

1) Cell viability (%)=(test compound well reading−blank wellreading)/(DMSO solvent control well reading−blank well reading)×100%;

2) a curve was plotted with GraphPad Prism 5.0 and IC₅₀ was obtained.

Experimental Result and Conclusion

TABLE 6 In vitro cytological activity (IC₅₀, nM) of compounds of thepresent invention Test compounds Ba/F3 SLC34A2/ROS1-G2032R Compound 32.3 Compound 11 4.2 Compound 13 6.7

It can be seen from Table 6 that the compounds of the present inventioncan effectively inhibit the proliferation of cells such as Ba/F3SLC34A2/ROS1-G2032R, indicating that the compounds of the presentinvention have the potential of clinical application in treatingdrug-resistant cancerous diseases caused by ROS gene mutation.

Experimental Example 3: Efficacy Study Experiment in Compound of thePresent Invention on BaF3-LMNA-NTRK1-G595R Stable Cell StrainSubcutaneous Xenograft Model Experimental Materials

The test compounds were compounds in the examples above.

Positive control: compound LOXO-195 with a structure shown as follows,prepared according to a prior art method (see patent applicationWO2011146336):

Experimental Method

1.1 Cell Inoculation

The RPMI 1640 serum-free medium resuspended BaF3 LMNA-NTRK1-G595R stablecell strains were inoculated subcutaneously in the right scapular regionof mice (NOD-SCID) at 1×10⁶ cells/mouse (0.1 mL/mouse).

1.2 Administration in Groups

When the average tumor volume was around 500 mm³, the groups wererandomized into vehicle group, Compound 3 group (10 mg/kg, 5 mg/kg, 3mg/kg, 1 mg/kg, bid) and LOXO-195 group (30 mg/kg, 10 mg/kg, 3 mg/kg,bid).

For the detailed administration method, administration dosage andadministration route, see Table 7-1 and Table 7-2.

TABLE 7-1 Efficacy experiment 1 inBaF3 LMNA-NTRK1-G595R model Admin-Admin- Administered Dose istration istration Group n group (mg/Kg) routetime 1 6 Vehicle group — — — 2 6 LOXO-195 group 3 p.o., bid 7 days 3 6LOXO-195 group 10 p.o., bid 7 days 4 6 Compound 3 3 p.o., bid 7 daysNote: n was the number of animals; the drug was administered on the dayof grouping.

TABLE 7-2 Efficacy experiment 2 in BaF3 LMNA-NTRK1-G595R model Admin-Admin- Administered Dose istration istration Group n group (mg/Kg) routetime 5 6 LOXO-195 group 30 p.o., bid 7 days 6 6 Compound 3 1 p.o., bid 7days 7 6 Compound 3 5 p.o., bid 7 days 8 6 Compound 3 10 p.o., bid 7days Note: n was the number of animals; the drug was administered on theday of grouping.

1.3 Experimental Evaluation Index

1) Tumor growth inhibition ratio TGI (%)

TGI (%)=(1−T/C)×100%

2) Tumor volume ratio T/C (%) (treatment group/control group)

T/C (%)=T _(RTV) /C _(RTV)×100%

*RTV=V_(t)/V₀, where V_(t) was the tumor volume on day t after grouping,V₀ was the tumor volume on the day of grouping;

T_(RTV): average relative tumor volume of the administered group;C_(RT)V: average relative tumor volume of the solvent control group.

3) The efficacy evaluation standard was as follows: ineffective when T/C(%) was more than 40%, and effective when the T/C (%) was less than orequal to 40%; effective when TGI was more than or equal to 60%.

Experimental Result and Conclusion

TABLE 8-1 Inhibition of Experiment 1 on BaF3-LMNA-NTRK1- G595R stablecell strain xenograft tumor Group Dosage (mg/Kg) T/C (%) TGI (%) 1(Vehicle group) — — 2 (LOXO-195 group) 3, bid 93.6 6.4 3 (LOXO-195group) 10, bid  63.1 36.9 4 (Compound 3 group) 3, bid 73.2 26.8

TABLE 8-2 Inhibition of Experiment 2 on BaF3-LMNA-NTRK1- G595R stablecell strain xenograft tumor Group Dosage (mg/Kg) T/C (%) TGI (%) 5(LOXO-195 group) 30, bid 0 100 6 (Compound 3 group)  1, bid 53.7 46.3 7(Compound 3 group)  5, bid 0 100 8 (Compound 3 group) 10, bid 0 100

Experimental data show that oral administration of an effective amountof compound of the present invention can obviously inhibit a cell lineBaF3-LMNA-NTRK1-G595R tumor efficacy model containing NTRK fusion genes,which means that the compound of the present invention shows good tumorinhibition effect on tumors with NTRK fusion gene mutation, and has agood clinical application prospect.

1. A compound of general formula (I) or a pharmaceutically acceptablesalt, ester or stereoisomer thereof,

wherein: M¹, M², M³, M⁴, M⁵, M⁶ and M⁷ are each independently selectedfrom C, C(R¹) and N; X¹, X², X³ and L¹, when present, are eachindependently selected from —C(R⁵)(R⁶)—, —N(R⁴)—, —O—, —S—, —S(O)— and—S(O)₂—; ring A is selected from 3-10 membered cycloalkyl, 3-10 memberedheterocyclyl, 6-8 membered monoaryl, 8-10 membered fused aryl, 5-10membered monoheteroaryl and 8-10 membered fused heteroaryl; ring B isabsent or selected from 3-10 membered cycloalkyl and 3-10 memberedheterocyclyl, and when ring B is absent, X² and X³ are directlyconnected by a chemical bond; R¹, R², R³, R⁵ and R⁶, when present, areeach independently selected from hydrogen, halogen, nitro, cyano and thefollowing groups optionally substituted with one or more Q¹: C₁₋₆ alkyl,C₂₋₆ alkenyl, C₂₋₆ alkynyl, —OR^(a), —SR^(a), —N^(a)R^(b), —C(O)R^(a),—C(O)OR^(a), —OC(O)R^(a), —OC(O)OR^(a), —OC(O)NR^(a)R^(b),—C(O)NR^(a)R^(b), —NR^(a)C(O)R^(b), —NR^(a)C(O)OR^(b),—NR^(a)C(O)NR^(a)R^(b), —S(O)R^(a), —S(O)OR^(a), —OS(O)R^(a),—OS(O)OR^(a), —OS(O)NR^(a)R^(b), —S(O)NR^(a)R^(b), —NR^(a)S(O)R^(b),—NR^(a)S(O)OR^(b), —NR^(a)S(O)NR^(a)R^(b), —S(O)₂R^(a), —S(O)₂OR^(a),—OS(O)₂R^(a), —OS(O)₂OR^(a), —OS(O)₂NR^(a)R^(b), —S(O)₂NR^(a)R^(b),—NR^(a)S(O)₂R^(b), —NR^(a)S(O)₂OR^(b), —NR^(a)S(O)₂NR^(a)R^(b), 3-10membered cycloalkyl, 3-10 membered heterocyclyl, 6-10 membered aryl and5-10 membered heteroaryl; Q¹, when present, is independently selectedfrom hydroxy, amino, halogen, nitro, cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy,C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino, halo C₁₋₆ alkyl, hydroxy C₁₋₆alkyl, amino C₁₋₆ alkyl, halo C₁₋₆ alkoxy, 3-10 membered cycloalkyl,3-10 membered heterocyclyl, 6-10 membered aryl and 5-10 memberedheteroaryl; R⁴, when present, is independently selected from hydrogenand the following groups optionally substituted with one or more Q²:C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, —C(O)R^(a), —C(O)OR^(a),—C(O)NR^(a)R^(b), —S(O)R^(a), —S(O)OR^(a), —S(O)NR^(a)R^(b),—S(O)₂R^(a), —S(O)₂OR^(a), —S(O)₂NR^(a)R^(b), 3-10 membered cycloalkyl,3-10 membered heterocyclyl, 6-10 membered aryl and 5-10 memberedheteroaryl; Q², when present, is independently selected from hydroxy,amino, halogen, nitro, cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ alkylamino,di(C₁₋₆ alkyl)amino, halo C₁₋₆ alkyl, hydroxy C₁₋₆ alkyl, amino C₁₋₆alkyl, halo C₁₋₆ alkoxy, 3-10 membered cycloalkyl, 3-10 memberedheterocyclyl, 6-10 membered aryl and 5-10 membered heteroaryl; or X¹,together with L¹, forms 3-10 membered cycloalkyl, 3-10 memberedheterocyclyl or 5-6 membered monoheteroaryl; and/or L¹, together withsome ring atoms of ring A, forms 3-10 membered cycloalkyl or 3-10membered heterocyclyl; and/or X², together with some ring atoms of ringA, forms 3-10 membered cycloalkyl or 3-10 membered heterocyclyl; the3-10 membered cycloalkyl, 3-10 membered heterocyclyl and 5-6 memberedmonoheteroaryl are optionally each independently substituted with asubstituent selected from: R⁴, R⁵, R⁶, halogen, amino, hydroxy, nitro,cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino,halo C₁₋₆ alkyl, hydroxy C₁₋₆ alkyl, amino C₁₋₆ alkyl and halo C₁₋₆alkoxy; R^(a) and R^(b), when present, are each independently selectedfrom hydrogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ alkylamino, di(C₁₋₆alkyl)amino, halo C₁₋₆ alkyl, hydroxy C₁₋₆ alkyl, amino C₁₋₆ alkyl, haloC₁₋₆ alkoxy, 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 6-10membered aryl and 5-10 membered heteroaryl; and m, n, p, q and t areeach independently selected from 0, 1, 2, 3, 4 and
 5. 2. The compound orthe pharmaceutically acceptable salt, ester or stereoisomer thereofaccording to claim 1, wherein: M¹, M², M³, M⁴, M⁵, M⁶ and M⁷ are eachindependently selected from C, C(R¹) and N, at least one of which is N;X¹, X², X³ and L¹, when present, are each independently selected from—C(R⁵)(R⁶)—, —N(R⁴)— and —O—; ring A is selected from 6-8 memberedmonoaryl and 5-8 membered monoheteroaryl; ring B is absent or selectedfrom 3-8 membered cycloalkyl and 3-8 membered heterocyclyl; R¹, R², R³,R⁵ and R⁶, when present, are each independently selected from hydrogen,halogen, nitro, cyano and the following groups optionally substitutedwith 1 to 3 Q¹: C₁₋₆ alkyl, —OR^(a), —NR^(a)R^(b), —C(O)R^(a),—C(O)OR^(a), —OC(O)R^(a), —OC(O)OR^(a), —OC(O)NR^(a)R^(b),—C(O)NR^(a)R^(b), —NR^(a)C(O)R^(b), —NR^(a)C(O)OR^(b),—NR^(a)C(O)NR^(a)R^(b), 3-8 membered cycloalkyl, 3-8 memberedheterocyclyl, 6-8 membered aryl and 5-8 membered heteroaryl; Q¹, whenpresent, is independently selected from hydroxy, amino, halogen, nitro,cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino,halo C₁₋₆ alkyl, hydroxy C₁₋₆ alkyl, amino C₁₋₆ alkyl and halo C₁₋₆alkoxy; R⁴, when present, is independently selected from hydrogen andthe following groups optionally substituted with 1 to 3 Q²: C₁₋₆ alkyl,—C(O)R^(a), —C(O)OR^(a), —C(O)NR^(a)R^(b), 3-8 membered cycloalkyl and3-8 membered heterocyclyl; Q², when present, is independently selectedfrom hydroxy, amino, halogen, nitro, cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy,C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino, halo C₁₋₆ alkyl, hydroxy C₁₋₆alkyl, amino C₁₋₆ alkyl, halo C₁₋₆ alkoxy, 3-8 membered cycloalkyl and3-8 membered heterocyclyl; or X¹, together with L¹, forms 3-8 memberedcycloalkyl, 3-8 membered heterocyclyl or 5-6 membered monoheteroaryl;and/or L¹, together with some ring atoms of ring A, forms 3-8 memberedcycloalkyl or 3-8 membered heterocyclyl; and/or X², together with somering atoms of ring A, forms 3-8 membered cycloalkyl or 3-8 memberedheterocyclyl; the 3-8 membered cycloalkyl, 3-8 membered heterocyclyl and5-6 membered monoheteroaryl are optionally each independentlysubstituted with a substituent selected from: R⁴, R⁵, R⁶, halogen,amino, hydroxy, nitro, cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ alkylamino,di(C₁₋₆ alkyl)amino, halo C₁₋₆ alkyl, hydroxy C₁₋₆ alkyl, amino C₁₋₆alkyl and halo C₁₋₆ alkoxy; R^(a) and R^(b), when present, are eachindependently selected from hydrogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆alkylamino, di(C₁₋₆ alkyl)amino, halo C₁₋₆ alkyl, hydroxy C₁₋₆ alkyl,amino C₁₋₆ alkyl, halo C₁₋₆ alkoxy, 3-8 membered cycloalkyl and 3-8membered heterocyclyl; and m, n, p, q and t are each independentlyselected from 0, 1, 2, 3, 4 and
 5. 3. The compound or thepharmaceutically acceptable salt, ester or stereoisomer thereofaccording to claim 2, wherein: M¹, M², M³, M⁴, M⁵, M⁶ and M⁷ are eachindependently selected from C, C(R¹) and N, at least one of which is N;X¹ and L¹ are each independently selected from —C(R⁵)(R⁶)— and —N(R⁴)—;X² and X³, when present, are each independently selected from—C(R⁵)(R⁶)—, —N(R⁴)— and —O—; ring A is selected from phenyl and 5-6membered monoheteroaryl; ring B is absent or selected from 3-6 memberedmonocycloalkyl and 3-6 membered monoheterocyclyl; R¹, R², R³, R⁵ and R⁶,when present, are each independently selected from hydrogen, halogen,nitro, cyano and the following groups optionally substituted with 1 to 3Q¹: C₁₋₆ alkyl, —OR^(a), —NR^(a)R^(b), —C(O)R^(a), —C(O)OR^(a),—C(O)NR^(a)R^(b), 3-6 membered cycloalkyl, 3-6 membered heterocyclyl,phenyl and 5-6 membered heteroaryl; Q¹, when present, is independentlyselected from hydroxy, amino, halogen, nitro, cyano, C₁₋₆ alkyl, C₁₋₆alkoxy, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino, halo C₁₋₆ alkyl, hydroxyC₁₋₆ alkyl, amino C₁₋₆ alkyl and halo C₁₋₆ alkoxy; R⁴, when present, isindependently selected from hydrogen and the following groups optionallysubstituted with 1 to 3 Q²: C₁₋₆ alkyl, —C(O)R^(a), 3-6 memberedcycloalkyl and 3-6 membered heterocyclyl; Q², when present, is selectedfrom hydroxy, amino, halogen, nitro, cyano, C₁₋₆ alkyl, C₁₋₆ alkoxy,C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino, halo C₁₋₆ alkyl, hydroxy C₁₋₆alkyl, amino C₁₋₆ alkyl, halo C₁₋₆ alkoxy, 3-6 membered cycloalkyl and3-6 membered heterocyclyl; or X¹, together with L¹, forms 3-6 memberedmonoheterocyclyl or 5-6 membered monoheteroaryl; and/or L¹, togetherwith some ring atoms of ring A, forms 3-6 membered monocycloalkyl or 3-6membered monoheterocyclyl; and/or X², together with some ring atoms ofring A, forms 3-6 membered monocycloalkyl and 3-6 memberedmonoheterocyclyl; the 3-6 membered monoheterocyclyl, 5-6 memberedmonoheteroaryl and 3-6 membered monocycloalkyl are optionally eachindependently substituted with a substituent selected from: R⁴, R⁵, R⁶,halogen, amino, hydroxy, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ alkylamino,di(C₁₋₆ alkyl)amino, halo C₁₋₆ alkyl, hydroxy C₁₋₆ alkyl, amino C₁₋₆alkyl and halo C₁₋₆ alkoxy; R^(a) and R^(b), when present, are eachindependently selected from hydrogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆alkylamino, di(C₁₋₆ alkyl)amino, halo C₁₋₆ alkyl, hydroxy C₁₋₆ alkyl,amino C₁₋₆ alkyl, halo C₁₋₆ alkoxy, 3-6 membered cycloalkyl and 3-6membered heterocyclyl; m is 1, 2 or 3; n, p and q are each independentlyselected from 0, 1, 2 and 3; and t is 0 or
 1. 4. The compound or thepharmaceutically acceptable salt, ester or stereoisomer thereofaccording to any one of claims 1-3, wherein the compound is of thestructure shown as general formula (II),

wherein M², M⁵ and M⁶ are each independently selected from CH and N. 5.The compound or the pharmaceutically acceptable salt, ester orstereoisomer thereof according to any one of claims 1-4, wherein thecompound is of the structure shown as general formula (III),

wherein: ring C is selected from 3-6 membered saturated monoheterocyclyland 5-6 membered nitrogen-containing monoheteroaryl, preferably 5-6membered saturated monoheterocyclyl; the 3-6 membered saturatedmonoheterocyclyl and 5-6 membered nitrogen-containing monoheteroaryl areoptionally each independently substituted with a substituent selectedfrom: R⁴, R⁵, R⁶, halogen, amino, hydroxy, C₁₋₄ alkyl, C₁₋₄ alkoxy, C₁₋₄alkylamino, di(C₁₋₄ alkyl)amino, halo C₁₋₄ alkyl, hydroxy C₁₋₄ alkyl,amino C₁₋₄ alkyl and halo C₁₋₄ alkoxy; —(X²)_(p)— is selected from—C(R⁵)(R⁶)—, —C(R⁵)(R⁶)—C(R⁵)(R⁶)—, —N(R⁴)—C(R⁵)(R⁶)—, —O—C(R⁵)(R⁶)—,—C(R⁵)(R⁶)—N(R⁴)— and —C(R⁵)(R⁶)—O—, and the left chemical bond thereofis connected to ring A and the right chemical bond thereof is connectedto X³; —(X³)_(q)— is selected from —C(R⁵)(R⁶)—, —C(R⁵)(R⁶)—C(R⁵)(R⁶)—,—N(R⁴)—C(R⁵)(R⁶)—, —O—C(R⁵)(R⁶)—, —C(R⁵)(R⁶)—N(R⁴)— and —C(R⁵)(R⁶)—O—,and the left chemical bond thereof is connected to X² and the rightchemical bond thereof is connected to carbonyl; ring A is selected fromphenyl and 5-6 membered monoheteroaryl, preferably 5-6 memberednitrogen-containing heteroaryl; R², when present, is independentlyselected from hydrogen, halogen and the following groups optionallysubstituted with 1 to 3 Q¹: C₁₋₄ alkyl, —OR^(a) and —NR^(a)R^(b); Q¹,when present, is independently selected from hydroxy, amino, halogen,nitro, cyano, C₁₋₄ alkyl, C₁₋₄ alkoxy, C₁₋₄ alkylamino, di(C₁₋₄alkyl)amino, halo C₁₋₄ alkyl, hydroxy C₁₋₄ alkyl, amino C₁₋₄ alkyl andhalo C₁₋₄ alkoxy; R^(a) and R^(b), when present, are each independentlyselected from hydrogen and C₁₋₄ alkyl; R⁵ and R⁶, when present, are eachindependently selected from hydrogen, halogen, hydroxy, amino, C₁₋₄alkyl and C₁₋₄ alkoxy; R⁴, when present, is independently selected fromhydrogen and C₁₋₄ alkyl optionally substituted with 1 to 2 Q²; Q², whenpresent, is independently selected from hydroxy, amino, halogen and C₁₋₄alkoxy; and n is 0, 1 or
 2. 6. The compound or the pharmaceuticallyacceptable salt, ester or stereoisomer thereof according to any one ofclaims 1-5, wherein the compound is of the structure shown as generalformula (IV),

wherein each optional substituent on ring C is independently selectedfrom R⁴, R⁵, R⁶, halogen, amino, hydroxy, nitro, cyano, C₁₋₆ alkyl, C₁₋₆alkoxy, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino, halo C₁₋₆ alkyl, hydroxyC₁₋₆ alkyl, amino C₁₋₆ alkyl and halo C₁₋₆ alkoxy.
 7. The compound orthe pharmaceutically acceptable salt, ester or stereoisomer thereofaccording to any one of claims 1-6, wherein: ring C is selected fromaziridinyl, azetidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,piperidinyl, morpholinyl, piperazinyl, hexahydropyridazinyl,hexahydropyrimidinyl, pyrrolyl, imidazolyl, pyrazolyl, pyridinyl orpyrimidinyl; each of which is optionally substituted with a substituentselected from: fluorine, chlorine, bromine, iodine, hydroxy, amino,methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, methoxy,ethoxy, propoxy, isopropoxy, methylamino, dimethylamino, ethylamino,diethylamino, trifluoromethyl and trifluoromethoxy; ring A is selectedfrom phenyl, pyrrolyl, imidazolyl, pyrazolyl, furyl, thienyl, thiazolyl,oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, 1,2,4-triazolyl,1,2,3-triazolyl, 1,3,4-oxadiazolyl, 1,2,4-oxadiazolyl,1,3,4-thiadiazolyl, 1,2,4-thiadiazolyl, pyridinyl, pyrimidinyl,pyridazinyl, 1,3,5-triazinyl and tetrazinyl; R², when present, isindependently selected from hydrogen, fluorine, chlorine, bromine,iodine, hydroxy, amino, methyl, ethyl, propyl, isopropyl, butyl,isobutyl, sec-butyl, methylamino, dimethylamino, ethylamino,diethylamino, methoxy, ethoxy, propoxy, isopropoxy, trifluoromethyl andtrifluoromethoxy; R⁵ and R⁶, when present, are each independentlyselected from hydrogen, fluorine, chlorine, bromine, iodine, hydroxy,amino, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl,methoxy, ethoxy, propoxy and isopropoxy; and R⁴, when present, isindependently selected from hydrogen, methyl, ethyl, propyl, isopropyl,butyl, isobutyl and sec-butyl.
 8. The compound or the pharmaceuticallyacceptable salt, ester or stereoisomer thereof according to any one ofclaims 1-7, wherein: ring A is selected from

and preferably the wavy line marked with “1” represents the connectionpoint of ring A to L¹ and the wavy line marked with “2” represents theconnection point of ring A to X²; —(X²)_(p)— is selected from—C(R⁵)(R⁶)—, —C(R⁵)(R⁶)—C(R⁵)(R⁶)—, —N(R⁴)—C(R⁵)(R⁶)— and —O—C(R⁵)(R⁶)—,and preferably the left chemical bond thereof is connected to ring A andthe right chemical bond thereof is connected to X³; and —(X³)_(q)— isselected from —C(R⁵)(R⁶)—, —C(R⁵)(R⁶)—N(R⁴)— and —C(R⁵)(R⁶)—O—, andpreferably the left chemical bond thereof is connected to X² and theright chemical bond thereof is connected to carbonyl.
 9. The compound orthe pharmaceutically acceptable salt, ester or stereoisomer thereofaccording to any one of claims 1-8, wherein the compound is selectedfrom the following compounds: No. Compound Compound 1

Compound 2

Compound 2-1

Compound 3

Compound 3-1

Compound 4

Compound 5

Compound 6

Compound 7

Compound 8

Compound 9

Compound 10

Compound 11

Compound 11-1

wherein * represents that a carbon atom is a chiral carbon atom having asingle configuration (R configuration or S configuration).
 10. Thecompound or the pharmaceutically acceptable salt, ester or stereoisomerthereof according to claim 9, wherein the compound is selected from: No.Compound Compound 2′

Compound 3′

Compound 11′


11. The compound or the pharmaceutically acceptable salt, ester orstereoisomer thereof according to any one of claims 1-4, wherein thecompound is of the structure shown as general formula (V),


12. The compound or the pharmaceutically acceptable salt, ester orstereoisomer thereof according to any one of claims 1-4, wherein: X¹ is—N(R⁴)—; L¹ is —C(R⁵)(R⁶)—; ring A is selected from phenyl and 5-6membered nitrogen-containing heteroaryl; R², when present, isindependently selected from hydrogen, halogen and the following groupsoptionally substituted with 1 to 3 Q¹: C₁₋₄ alkyl, —OR^(a) and—NR^(a)R^(b); Q¹, when present, is independently selected from hydroxy,amino, halogen, nitro, cyano, C₁₋₄ alkyl, C₁₋₄ alkoxy, C₁₋₄ alkylamino,di(C₁₋₄ alkyl)amino, halo C₁₋₄ alkyl, hydroxy C₁₋₄ alkyl, amino C₁₋₄alkyl and halo C₁₋₄ alkoxy; R^(a) and R^(b), when present, are eachindependently selected from hydrogen and C₁₋₄ alkyl; R⁵ and R⁶, whenpresent, are each independently selected from hydrogen, halogen,hydroxy, amino, C₁₋₄ alkyl and C₁₋₄ alkoxy; R⁴, when present, isindependently selected from hydrogen and C₁₋₄ alkyl optionallysubstituted with 1 to 2 Q²; Q², when present, is independently selectedfrom hydroxy, amino, halogen and C₁₋₄ alkoxy; and n is 0, 1 or
 2. 13.The compound or the pharmaceutically acceptable salt, ester orstereoisomer thereof according to claim 12, wherein: ring A is selectedfrom phenyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl,isoxazolyl, thiazolyl, isothiazolyl, 1,2,4-triazolyl, 1,2,3-triazolyl,1,3,4-oxadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-thiadiazolyl,1,2,4-thiadiazolyl, pyridinyl, pyrimidinyl, pyridazinyl, 1,3,5-triazinyland tetrazinyl; R², when present, is independently selected fromhydrogen, fluorine, chlorine, bromine, iodine, hydroxy, amino, methyl,ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, methylamino,dimethylamino, ethylamino, diethylamino, methoxy, ethoxy, propoxy,isopropoxy, trifluoromethyl and trifluoromethoxy; R⁵ and R⁶, whenpresent, are each independently selected from hydrogen, fluorine,chlorine, bromine, iodine, hydroxy, amino, methyl, ethyl, propyl,isopropyl, butyl, isobutyl, sec-butyl, methoxy, ethoxy, propoxy andisopropoxy; and R⁴, when present, is independently selected fromhydrogen, methyl, ethyl, propyl, isopropyl, butyl, isobutyl andsec-butyl.
 14. The compound or the pharmaceutically acceptable salt,ester or stereoisomer thereof according to claim 12, wherein: ring A isselected from

and preferably the wavy line marked with “1” represents the connectionpoint of ring A to L¹ and the wavy line marked with “2” represents theconnection point of ring A to X²; —(X²)_(p)— is selected from—C(R⁵)(R⁶)—, —C(R⁵)(R⁶)—C(R⁵)(R⁶)—, —N(R⁴)—C(R⁵)(R⁶)— and —O—C(R⁵)(R⁶)—,and preferably the left chemical bond thereof is connected to ring A andthe right chemical bond thereof is connected to X³; and —(X³)_(q)— isselected from —C(R⁵)(R⁶)—, —C(R⁵)(R⁶)—N(R⁴)— and —C(R⁵)(R⁶)—O—, andpreferably the left chemical bond thereof is connected to X² and theright chemical bond thereof is connected to carbonyl.
 15. The compoundor the pharmaceutically acceptable salt, ester or stereoisomer thereofaccording to any one of claims 12-14, wherein the compound is selectedfrom: No. Compound Compound 12

Compound 13

Compound 13-1

Compound 14

Compound 15

Compound 16

Compound 17


16. A pharmaceutical formulation comprising the compound or thepharmaceutically acceptable salt, ester or stereoisomer thereofaccording to any one of claims 1-15, and one or more pharmaceuticallyacceptable carriers and/or excipients.
 17. A pharmaceutical compositioncomprising the compound or the pharmaceutically acceptable salt, esteror stereoisomer thereof according to any one of claims 1-15, and one ormore second therapeutically active agents selected from: a drug for thetreatment of pain, which is selected from Nav1.7 channel modulators,opioid analgesics, non-steroidal anti-inflammatory drugs, sedatives,selective/non-selective cyclooxygenase inhibitors, antiepileptics,antidepressants, local anesthetics, 5-HT receptor blockers, 5-HTreceptor agonists, ergot alkaloids, β-receptor blockers, M receptorblockers, nitrates and vitamin K; a drug for the treatment of cancer,which is selected from mitotic inhibitors, alkylating agents,antimetabolites, antisense DNA or RNA, antitumor antibiotics, growthfactor inhibitors, signal transduction inhibitors, cell cycleinhibitors, enzyme inhibitors, retinoid receptor modulators, proteasomeinhibitors, topoisomerase inhibitors, biological response modifiers,hormones, angiogenesis inhibitors, cell growth inhibitors, targetedantibodies, HMG-CoA reductase inhibitors and prenyl-protein transferaseinhibitors; a drug for the treatment of neurodegenerative diseases,which is selected from dopamine-mimetics, dopamine receptor agonists,agents affecting dopamine metabolism, NMDA receptor antagonists,adenosine A_(2A) receptor inhibitors, agents affecting DA release andreuptake, central anticholinergics, cholinesterase inhibitors, 5-HTagonists, α2 adrenergic receptor antagonists, antidepressants,cholinergic receptor agonists, β/γ secretase inhibitors, H3 receptorantagonists and antioxidant agents; a drug for the treatment ofautoimmune diseases, which is selected from antirheumatic drugs,non-steroidal anti-inflammatory drugs, glucocorticoid drugs, TNFantagonists, cyclophosphamide, mycophenolate mofetil and cyclosporine,which is used for alleviating the disease; and a drug for the treatmentof inflammation, which is selected from steroidal anti-inflammatorydrugs and non-steroidal anti-inflammatory drugs.
 18. Use of the compoundor the pharmaceutically acceptable salt, ester or stereoisomer thereofaccording to any one of claims 1-15 in the preparation of a medicationfor the treatment and/or prevention of a disease or related conditionmediated by one or more tyrosine kinases of TRK, ALK and/or ROS1,wherein the disease or related condition is selected from one or more ofpain, cancers, inflammation, neurodegenerative diseases and autoimmunediseases.
 19. Use of the compound or the pharmaceutically acceptablesalt, ester or stereoisomer thereof according to claim 18, in thepreparation of a medication for the treatment and/or prevention of adisease or related a condition mediated by one or more tyrosine kinasesof TRK, ALK and/or ROS1, wherein the disease is, for example, a cancerthat is at least partially resistant to a therapeutically active agenttargeting TRK, ALK and/or ROS1.
 20. The use of claim 19, wherein thedisease mediated by one or more tyrosine kinases of TRK, ALK and/or ROS1is a cancer, wherein the cancer is selected from lung cancer, coloncancer, rectal cancer, prostate cancer, breast cancer, liver cancer,gallbladder cancer, cholangiocarcinoma, leukemia, melanoma, lymphoma,thyroid cancer, multiple myeloma, soft tissue sarcoma, ovarian cancer,cervical cancer, fallopian tube carcinoma, renal cell carcinoma, gastriccancer, gastrointestinal stromal tumor, bone cancer, basal cellcarcinoma, peritoneal cancer, dermatofibroma, pancreatic cancer,esophageal cancer, glioblastoma, head and neck cancer, inflammatorymyofibroblast tumor, anaplastic large cell lymphoma and neuroblastoma;and preferably the lung cancer is selected from small cell lung cancerand non-small cell lung cancer.
 21. A method for preparing the compoundof general formula (I) or the pharmaceutically acceptable salt, ester orstereoisomer thereof,

wherein the substituents and variables are defined as any one of claims1-15, and the method comprises the steps as follows: intermediate 1 andintermediate 2 are subjected to a plurality of reactions such assubstitution reaction, deprotection reaction, acylation reaction (usingan acylation reagent) and the like in the presence of an organic solventand a catalyst at a proper temperature to obtain the compound of thegeneral formula (I).